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Abstract Background Combination therapy including an aminoglycoside antibiotic and a cell-wall active agent is considered the most suitable option to treat invasive infections with methicillin-resistant Staphylococcus aureus (MRSA). Dual drug therapy enhances the effectiveness of treatment and reduces the risk of resistance development. This study aims to elucidate the phenotypic and molecular resistance to aminoglycosides and methicillin, and the molecular epidemiologic characteristics of S. aureus in Ardabil northwest Iran. Methods Totally, 118 S. aureus isolates collected from clinical specimens were investigated. Identification was performed using standard microbiological and molecular approaches. Aminoglycoside and methicillin resistance were evaluated using the disk diffusion assay, and the minimum inhibitory concentrations (MICs) of aminoglycosides were determined via the agar dilution method. The mecA gene encoding methicillin resistance and aminoglycoside modifying enzymes (AMEs) genes were detected using PCR. Molecular epidemiologic features of the isolates were determined using staphylococcal cassette chromosome mec (SCCmec) typing spa typing and ERIC-PCR assays. Results Of the isolates, 42.4% (n = 50) and 57.6% (n = 68) were identified as MRSA and MSSA, respectively. All MRSA isolates were mecA-positive. Among MRSA isolates, SCCmec type IVa (17; 34%) was predominant, followed by types IVc, V, III, II, and I. Resistance rates to gentamicin, kanamycin, tobramycin, and amikacin were 16.1%, 17.8%, 8.5%, and 8.5%, respectively. Overall, the aminoglycoside resistance and most non-aminoglycoside antibiotics were significantly higher in MRSA versus MSSA isolates. The prevalence of AME genes was as follows: aac(6’)-Ie-aph(2’’) (30; 76.9%), aph(2’’)-Ib (22; 56.4%), and ant(4’)-Ia (14; 35.9%). About 60% of aminoglycoside-resistant isolates harbored ≥ 2 AME genes. The t030 type was the most common spa type identified. The ERIC-PCR profiles categorized the isolates into 19 unique ERIC types. Conclusions This study reveals high aminoglycoside and methicillin resistance in S. aureus isolates from Ardabil hospitals. Predominant SCCmec type IVa and spa type t030 indicate specific molecular patterns. These findings highlight the need for continuous surveillance and targeted treatment strategies for MRSA infections. Clinical trial number Not applicable.

Background: Characteristics of non-clinical strains of methicillin-resistant Staphylococcus aureus (MRSA) especially from fishery environment are poorly understood. This research, in addition to comprehensive characterisation, sought to delineate the genetic relatedness between the MRSA strains originating from clinical as well as non-clinical settings. Out of 39 methicillin-resistant staphylococcal isolates from 197 fish samples, 6 (Three each of methicillin-resistant S. haemolyticus (MRSH) and MRSA) with distinct resistance profiles were selected for whole-genome sequencing. Using respective bioinformatics tools, MRSA genomes were comprehensively characterized for resistome, virulomes, molecular epidemiology and phylogenetic analysis. Simultaneously, MRSH genomes were specifically examined to characterize antimicrobial resistance genes (ARGs), owing to the fact that MRSH is often recognized as a reservoir for resistance determinants. Results: Three MRSA clones identified in this study include ST672-IVd/t13599 (sequence type-SCCmec type/spa type), ST88-V/t2526, and ST672-IVa/t1309. Though, the isolates were phenotypically vancomycin-sensitive, five of the six genomes carried vancomycin resistance genes including the VanT (VanG cluster) or VanY (VanM cluster). Among the three MRSA, only one harbored the gene encoding Panton-Valentine Leukocidin (PVL) toxin, while staphylococcal enterotoxin (SEs) genes such as sea and seb, associated with staphylococcal food poisoning were identified in two other MRSA. Genomes of MRSH carried a composite of type V staphylococcal cassette chromosome mec (SCCmec) elements (5C2 & 5). This finding may be explained by the inversion and recombination events that may facilitate the integration of type V elements to the SCC elements of S. aureus with a methicillin-susceptible phenotype. Phylogenetically, MRSA from a non-clinical setting displayed a considerable relatedness to that from clinical settings. Conclusion: This study highlights the genetic diversity and resistance profiles of MRSA and MRSH, with non-clinical MRSA showing notable relatedness to clinical strains. Future research should explore resistance gene transfer mechanisms and environmental reservoirs to better manage MRSA spread. [ABSTRACT FROM AUTHOR]

The emergence and spread of Staphylococcus aureus, which is resistant to quinolone antibacterial agents, has made it difficult to treat infectious diseases. Accordingly, this study examined the molecular epidemiological characteristics of quinolone-resistant S. aureus (QRSA) to obtain helpful data for treatment. Mutations in mecA and SCCmec typing, gyrA, and gyrB genes were investigated for QRSA strains isolated from the blood culture specimens at a general hospital in Daejeon Metropolitan City. The ciprofloxacin-resistant strains in SCCmec typing were II (44 strains, 73%), IVa (five strains, 8%), III, and V (one strain, 2%); the non-typeable strains (11 strains, 18%), and levofloxacin (LVX) and moxifloxacin (MXF) strains were II (44 strains, 73%), IVa (five strains, 8%), III, and V (one strain, 2%); the non-typeable strains were 10 (17%). In both gyrA and gyrB regions, there were 58 mutations, or 96.7%. In LVX, there were 56 mutations or 93.3%, and in MXF, there were 57 mutations or 95%. Twelve mutations, six mutations each in gyrA and gyrB, were identified for the QRSA strain. The resistance rate for the quinolone antibiotics of QRSA studied was approximately 98%, and 12 mutations, six each in gyrA and gyrB, were identified in the QRSA strain. Therefore, the rational use of antibiotics needs to be improved.

Abstract Mastitis is considered one of the most widespread infectious disease of cattle and buffaloes, affecting dairy herds. The current study aimed to characterize the Staphylococcus aureus isolates recovered from subclinical mastitis animals in Pothohar region of the country. A total of 278 milk samples from 17 different dairy farms around two districts of the Pothohar region, Islamabad and Rawalpindi, were collected and screened for sub clinical mastitis using California Mastitis Test. Positive milk samples were processed for isolation of Staphylococcus aureus using mannitol salt agar. The recovered isolates were analyzed for their antimicrobial susceptibility and virulence genes using disc diffusion and PCR respectively. 62.2% samples were positive for subclinical mastitis and in total 70 Staphylococcus aureus isolates were recovered. 21% of these isolates were determined to be methicillin resistant, carrying the mecA gene. S. aureus isolates recovered during the study were resistant to all first line therapeutic antibiotics and in total 52% isolates were multidrug resistant. SCCmec typing revealed MRSA SCCmec types IV and V, indicating potential community-acquired MRSA (CA-MRSA) transmission. Virulence profiling revealed high prevalence of key genes associated with adhesion, toxin production, and immune evasion, such as hla, hlb, clfA, clfB and cap5. Furthermore, the Panton-Valentine leukocidin (PVL) toxin, that is often associated with recurrent skin and soft tissue infections, was present in 5.7% of isolates. In conclusion, the increased prevalence of MRSA in bovine mastitis is highlighted by this study, which also reveals a variety of virulence factors in S. aureus and emphasizes the significance of appropriate antibiotic therapy in combating this economically burdensome disease.

Abstract Methicillin-resistant Staphylococcus (MRS) has been associated with neonatal infections, with colonization of the anovaginal tract being the main source of vertical transmission. The COVID-19 pandemic has altered the frequency of antibiotic usage, potentially contributing to changes in the dynamics of bacterial agents colonizing humans. Here we determined MRS colonization rates among pregnant individuals attending a single maternity in Rio de Janeiro, Brazil before (January 2019–March 2020) and during (May 2020–March 2021) the COVID-19 pandemic. Anovaginal samples (n = 806 [521 samples before and 285 during the pandemic]) were streaked onto chromogenic media. Colonies were identified by MALDI-TOF MS. Detection of mecA gene and SCCmec typing were assessed by PCR and antimicrobial susceptibility testing was done according to CLSI guidelines. After the onset of the pandemic, MRS colonization rates increased significantly (p

Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) has been known as an infectious pathogen worldwide since 1960. The epidemiological distribution of MRSA may have shifted due to outbreaks reported from several nations, making it more challenging to differentiate among CA-MRSA and HA-MRSA. It is currently important to develop a strain-based explanation for HA and CA-MRSA due to its distinct epidemiology, genetic profile, antibiogram, and quantifiable features. The study aimed to distinguish CA and HA-MRSA by Staphylococcal Cassette Chromosome mec (SCCmec) typing. Materials and Method: The study involved a total of 381 S. aureus isolates, which were processed in the department of Microbiology, JSS Hospital, Mysore. All isolates were confirmed as MRSA, initially by disk diffusion method using cefoxitin 30µg and oxacillin 1μg disk and later by using PCR technique for the detection of mecA-gene. All mecA-gene positive samples were amplified for SCCmec typing by multiplex PCR for detection of SCCmec type I, II, III, IVa, IVb, IVc, IVd, V and XI respectively. Results: PCR confirmed a total of 66% isolates as mecA-positive MRSA. Multiplex PCR method revealed only 53% isolates were SCCmec-typeable. The mainstream of the isolates belonged to SCCmec type IV (53.48%) and type V (44.18%), followed by type III (9.30%), type II (3%) and type I (1.16%) respectively. The study also demonstrated the presence of multiple SCCmec types in 10.46% of MRSA isolates. Staphylococcal cassette chromosome recombinase (ccr) typing determined only 43% of isolates were typeable. Conclusion: The study found that hospital-associated SCCmec type IV and type V were the most circulating strains in our healthcare setting. The research identified a few MRSA isolates with diverse SCCmec types. The presence of CAMRSA infection in in-patients and HA-MRSA infection in out-patients were also recognised. The situation indicates the spreading of hospital-acquired strains into the community and vice versa, necessitating the molecular characterisation of MRSA isolates in order to administer the appropriate antibiotic medication. [ABSTRACT FROM AUTHOR]

Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) has been known as an infectious pathogen worldwide since 1960. The epidemiological distribution of MRSA may have shifted due to outbreaks reported from several nations, making it more challenging to differentiate among CA-MRSA and HA-MRSA. It is currently important to develop a strain-based explanation for HA and CA-MRSA due to its distinct epidemiology, genetic profile, antibiogram, and quantifiable features. The study aimed to distinguish CA and HA-MRSA by Staphylococcal Cassette Chromosome mec (SCCmec) typing. Materials and Method: The study involved a total of 381 S. aureus isolates, which were processed in the department of Microbiology, JSS Hospital, Mysore. All isolates were confirmed as MRSA, initially by disk diffusion method using cefoxitin 30µg and oxacillin 1μg disk and later by using PCR technique for the detection of mecA-gene. All mecA-gene positive samples were amplified for SCCmec typing by multiplex PCR for detection of SCCmec type I, II, III, IVa, IVb, IVc, IVd, V and XI respectively. Results: PCR confirmed a total of 66% isolates as mecA-positive MRSA. Multiplex PCR method revealed only 53% isolates were SCCmec-typeable. The mainstream of the isolates belonged to SCCmec type IV (53.48%) and type V (44.18%), followed by type III (9.30%), type II (3%) and type I (1.16%) respectively. The study also demonstrated the presence of multiple SCCmec types in 10.46% of MRSA isolates. Staphylococcal cassette chromosome recombinase (ccr) typing determined only 43% of isolates were typeable. Conclusion: The study found that hospital-associated SCCmec type IV and type V were the most circulating strains in our healthcare setting. The research identified a few MRSA isolates with diverse SCCmec types. The presence of CA-MRSA infection in in-patients and HA-MRSA infection in out-patients were also recognised. The situation indicates the spreading of hospital-acquired strains into the community and vice versa, necessitating the molecular characterisation of MRSA isolates in order to administer the appropriate antibiotic medication.

Background: Methicillin-resistant Staphylococcus aureus (MRSA), is considered a potential and aggressive nosocomial pathogen. It accounts for 50% of S. aureus isolates in tertiary hospitals in Iran, however, there is no sufficient evolutionary and epidemiological investigation about this medically important bacterium. We aimed to study the lineage and evolution of MRSA in Northwest Iran during 2021–2022 based on the obtained phenotypic and genotypic characteristics. Materials and methods: Seventy-two non-duplicate MRSA isolates were collected from 3 referral hospitals in Tabriz, Ardebil, and Urmia cities. The antimicrobial susceptibility patterns were determined by disk diffusion test and micro broth dilution methods. Thereafter 4 virulence genes (eta, etb, pvl, tst) and 5 types of staphylococcal cassette chromosome mec (SCCmec) were detected by PCR. In the final step, representative isolates were selected to be studied by Multilocus sequence typing (MLST). Results: The highest resistance was observed to erythromycin and clindamycin at a rate of 76.4%, followed by ciprofloxacin (61.1%), gentamicin (54.2%), rifampin (38.9%), and co-trimoxazole (27.8%). All isolates were susceptible to vancomycin. The virulence genes of etb, pvl, tst, and eta were detected in 50%, 29.2%, 21.8%, and 13.9% of isolates, respectively. SCCmec types III and I were the most prevalent types, followed by types IV, II, and V. MLST analysis revealed 6 sequence types: ST6854, ST5282, ST127, ST7804, ST1607, and ST7784. Two MLST-based clonal complexes (CC8, and CC97) were identified as well. Conclusion: The ST numbers were non-repetitive. CC8 as a pandemic clone and an individual lineage and clinically significant clade was reported as the most prevalent clonal complex. It is essential periodic evaluations of antibiotic susceptibility patterns and study the evolutionary characteristics of medical-challenging microorganisms in particular MRSA to effectively treat and restrict the outbreaks. [ABSTRACT FROM AUTHOR]

Macau, recognized as a global tourism hub and the world's most densely populated region, provides a unique environment conducive to methicillin-resistant Staphylococcus aureus (MRSA) transmission in healthcare and community settings, posing a significant public health concern both locally and globally. The epidemiology and molecular characteristics of MRSA in the distinct city of Macau remain largely unelucidated. This five-year longitudinal study (2017–2022) examined the local prevalence and molecular typing of MRSA in Macau, with future MRSA type distribution predicted through ARIMA modeling. We subsequently analyzed the epidemiological characteristics of MRSA, including specimen source, clinical department, collection year, season, patient age, sex, and the annual number of tourists. Comprehensive antibiotic resistance profiles of the strains were also assessed. Of 504 clinically isolated S. aureus strains, 183 (36.3%) were identified as MRSA by the cefoxitin disk diffusion method and validated through multi-locus sequence typing (MLST). The MRSA detection rate showed an upward trend, increasing from 30.1% in 2017 to 45.7% in 2022. SCCmec type IV was predominant (28.9%), followed by types II (25.4%), III (22.1%), and V (22.1%). The primary sources of MRSA isolates were sputum (39.2%) and secretions (25.6%). Older age emerged as a risk factor for MRSA infection, whereas no significant associations were found with seasonal variations, gender, or the annual number of tourists. Despite displaying universal resistance to cefoxitin, oxacillin, and benzylpenicillin, MRSA isolates in Macau remained fully sensitive to vancomycin, tigecycline, quinupristin, nitrofurantoin, and linezolid. Continuous surveillance and analysis of MRSA distribution in Macau could provide invaluable insights for the effective management of MRSA prevention and control measures within healthcare settings. [ABSTRACT FROM AUTHOR]

Objective: To determine the antibiotic resistance profile (ARP) of Staphylococcus (S.) aureus isolates and molecular typing of the methicillin-resistant S. aureus (MRSA) isolates from Tuanku Mizan Armed Forces Hospital (TMAFH), Kuala Lumpur. Methods: The ARP and presence of the pvl gene were determined for 209 S. aureus isolates from clinical specimens. Of these, 123 were methicillin-susceptible S. aureus (MSSA) isolates and 86 were MRSA isolates. All MRSA isolates were characterized using SCCmec typing and spa typing. Descriptive analysis was performed to compare the demographic data with the phenotypic and genotypic variables of the S. aureus isolates. Results: No vancomycin-intermediate and -resistant S. aureus (VISA and VRSA, respectively) were detected among the study isolates. The MSSA isolates showed low resistance rates to all tested antibiotics, were commonly invasive (28/42, 66.7%), and mostly harboured pvl (35/42, 83.3%). Meanwhile, MRSA isolates showed high resistance to penicillin (86/86, 100%), ampicillin (86/86, 100%), sulbactam/ampicillin (86/86, 100%), cefuroxime (81/86, 94.19%), cefoperazone (76/86, 88.37%), azithromycin (56/86, 65.12%), and erythromycin (54/86, 62.79%). The majority of MRSA isolates were of SCCmec type IVh (65/86, 75.58%), spa type t032 (55/85, 63.95%), and grouped into spaCC-t022 (66/85, 77.65%). The t032 type was found to be associated with resistance traits to azithromycin and erythromycin (P

Background Methicillin-resistant Staphylococcus aureus (MRSA) is a widely recognized multidrug-resistant bacteria presenting a major therapeutic challenge to clinicians. Staphylococcus aureus possesses a number of pathogenicity factors that attribute to the severity of infections. This study was undertaken to investigate the common virulence genes in clinical isolates of Staphylococcus aureus , determine their antimicrobial susceptibility profile, and to characterize the staphylococcal cassette chromosome mec (SCCmec) types among MRSA in a tertiary care center. Materials and Methods A total of 133 clinical isolates were included in this study. Susceptibility to various antibiotics was determined by disc diffusion method. Methicillin resistance was screened using cefoxitin disc; m ecA and mecC genes were detected using polymerase chain reaction (PCR). PCR was done to detect 12 virulence factors such as hla , hlb , fnbA , fnbB , sea , seb , sec , icaA , clfA , tst , pvl, and eta. SCCmec typing was done by multiplex PCR. Results Of the 133 clinical isolates, 54 (40.6%) were MRSA. The most common virulence gene detected was hlb (61.6%), hla (39%), and fnbA (37%). SCCmec type I was the most predominant. Mortality rate of 6.7% was observed among patients with staphylococcal infections. Univariate analysis of mortality associated virulence genes did not reveal any significant association between virulence genes and mortality. Conclusion The distribution of virulence genes is similar in both MRSA and methicillin-sensitive Staphylococcus aureus. MRSA belongs to the SCCmec types I to IV. Possession of multiple virulence factors and multidrug resistance profile makes Staphylococcus aureus a formidable pathogen in clinical settings. [ABSTRACT FROM AUTHOR]

Objective: To determine the antibiotic resistance profile (ARP) of Staphylococcus (S.) aureus isolates and molecular typing of the methicillin-resistant S. aureus (MRSA) isolates from Tuanku Mizan Armed Forces Hospital (TMAFH), Kuala Lumpur. Methods: The ARP and presence of the pvl gene were determined for 209 S. aureus isolates from clinical specimens. Of these, 123 were methicillin-susceptible S. aureus (MSSA) isolates and 86 were MRSA isolates. All MRSA isolates were characterized using SCCmec typing and spa typing. Descriptive analysis was performed to compare the demographic data with the phenotypic and genotypic variables of the S. aureus isolates. Results: No vancomycin-intermediate and -resistant S. aureus (VISA and VRSA, respectively) were detected among the study isolates. The MSSA isolates showed low resistance rates to all tested antibiotics, were commonly invasive (28/42, 66.7%), and mostly harboured pvl (35/42, 83.3%). Meanwhile, MRSA isolates showed high resistance to penicillin (86/86, 100%), ampicillin (86/86, 100%), sulbactam/ampicillin (86/86, 100%), cefuroxime (81/86, 94.19%), cefoperazone (76/86, 88.37%), azithromycin (56/86, 65.12%), and erythromycin (54/86, 62.79%). The majority of MRSA isolates were of SCCmec type IVh (65/86, 75.58%), spa type t032 (55/85, 63.95%), and grouped into spaCC-t022 (66/85, 77.65%). The t032 type was found to be associated with resistance traits to azithromycin and erythromycin (P<0.05). We also found several spa types that are typically associated with hospital-, community-, and livestock-associated MRSA co-existing in our MRSA population. Conclusions: This study reflected the consistent absence of VISA and VRSA and corroborated the clonal shifting of MRSA isolates in the Malaysian MRSA isolates. [ABSTRACT FROM AUTHOR]

Macau, recognized as a global tourism hub and the world’s most densely populated region, provides a unique environment conducive to methicillin-resistant Staphylococcus aureus (MRSA) transmission in healthcare and community settings, posing a significant public health concern both locally and globally. The epidemiology and molecular characteristics of MRSA in the distinct city of Macau remain largely unelucidated. This five-year longitudinal study (2017–2022) examined the local prevalence and molecular typing of MRSA in Macau, with future MRSA type distribution predicted through ARIMA modeling. We subsequently analyzed the epidemiological characteristics of MRSA, including specimen source, clinical department, collection year, season, patient age, sex, and the annual number of tourists. Comprehensive antibiotic resistance profiles of the strains were also assessed. Of 504 clinically isolated S. aureus strains, 183 (36.3%) were identified as MRSA by the cefoxitin disk diffusion method and validated through multi-locus sequence typing (MLST). The MRSA detection rate showed an upward trend, increasing from 30.1% in 2017 to 45.7% in 2022. SCCmec type IV was predominant (28.9%), followed by types II (25.4%), III (22.1%), and V (22.1%). The primary sources of MRSA isolates were sputum (39.2%) and secretions (25.6%). Older age emerged as a risk factor for MRSA infection, whereas no significant associations were found with seasonal variations, gender, or the annual number of tourists. Despite displaying universal resistance to cefoxitin, oxacillin, and benzylpenicillin, MRSA isolates in Macau remained fully sensitive to vancomycin, tigecycline, quinupristin, nitrofurantoin, and linezolid. Continuous surveillance and analysis of MRSA distribution in Macau could provide invaluable insights for the effective management of MRSA prevention and control measures within healthcare settings.

The global emergence of methicillin-resistant Staphylococcus aureus (MRSA) that unsusceptible to a wide selection of antimicrobial agents and any newly introduced antimicrobial over the past decades has triggered more extensive holistic measures to put an end to this situation. Molecular surveillance of MRSA clones is important to understand their evolutionary dynamics for investigating outbreaks, propagating precautionary measures, as well as planning for appropriate treatment. This review includes peer-reviewed reports on the molecular characterisation of clinical Staphylococcus aureus isolates within Malaysian hospitals from year 2008 to 2020. This work highlights the molecular clones of hospital-acquired MRSA (HA-MRSA) and community-acquired MRSA (CA-MRSA) isolates from Malaysian hospitals, with description on their ever-changing pattern. Among HA-MRSA, the ST22-t032-SCCmec IV MRSA clone was reported to supplant the previous dominating clone, ST239-t037-SCCmec III. Meanwhile, ST30, ST772, ST6 and ST22 were repeatedly detected in CA-MRSA, however, none of the strains became predominant. Future in-depth study on molecular epidemiology of MRSA clone is essential for the investigation of the extent of the clonal shift, especially in Malaysia. [ABSTRACT FROM AUTHOR]

Multidrug-resistant Staphylococcus aureus strains have been commonly found in hospitals and communities causing wide ranges of infections among humans and animals. Typing of these strains is a key factor to reveal their clonal dissemination in different regions. We investigated the prevalence and dissemination of different clonal groups of S. aureus with resistance phenotype to multiple antibiotics in two sewage treatment plants (STPs) in Tehran, Iran over four sampling occasions. A total of 576 S. aureus were isolated from the inlet, sludge and outlet. Of these, 80 were identified as methicillin-resistant S. aureus (MRSA) and were further characterized using a combination of Phene Plate (PhP) typing, staphylococcal cassette chromosome mec (SCCmec), ccr types, prophage and antibiotic-resistant profiling. In all, eight common type (CT) and 13 single PhP type were identified in both STPs, with one major CT accounting for 38.8% of the MRSA strains. These strains belonged to three prophage patterns and five prophage types with SCCmec type III being the predominant type. Resistance to 11 out of the 17 antibiotics tested was significantly (P < 0.0059) higher among the MRSA isolates than methicillin-sensitive S. aureus (MSSA) strains. The persistence of the strains in samples collected from the outlet of both STPs was 31.9% for MRSA and 23.1% for MSSA. These data indicated that while the sewage treatment process, in general, is still useful for removing most MRSA populations, some strains with SCCmec type III may have a better ability to survive the STP process. HIGHLIGHTS Survival of MRSA in STPs has a major impact on public health if they enter surface waters.; Here we show that certain clones of MRSA carrying SCCmec type III have a better ability to survive treatment stages of the STPs.; The fact that these clones were resistant to up to 12 antibiotics suggests that these clones may also have a better ability to either gain or retain antibiotic-resistant genes during the STP process.;

Background: Staphylococcus aureus is often considered as a potential pathogen and resistant to a wide range of antibiotics. The pathogenicity of this bacterium is due to the presence of multiple virulence factors and the ability to form biofilm. SCCmec types I, II and III are mainly attributed to HA-MRSA, while SCCmec types IV and V have usually been reported in CA-MRSA infections. Methods and results: In this study, we performed a cross-sectional study to determine the antimicrobial resistance, adhesion and virulence factors, biofilm formation and SCCmec typing of clinical S. aureus isolates in Iran. S. aureus isolates were identified using microbiological standard methods and antibiotic susceptibility tests were performed as described by the Clinical and Laboratory Standards Institute (CLSI) guidelines. Inducible resistance phenotype and biofilm formation were determined using D-test and tissue culture plate methods, respectively. Multiplex-PCRs were performed to detect adhesion and virulence factors, antibiotic resistance genes, biofilm formation and SCCmec typing by specific primers. Among 143 clinical samples, 67.8% were identified as MRSA. All isolates were susceptible to vancomycin. The prevalence of cMLSB, iMLSB and MS phenotypes were 61.1%, 22.2% and 14.8%, respectively. The TCP method revealed that 71.3% of isolates were able to form biofilm. The predominant virulence and inducible resistance genes in both MRSA and MSSA isolates were related to sea and ermC respectively. SCCmec type III was the predominant type. Conclusions: Data show the high prevalence rates of virulence elements among S. aureus isolates, especially MRSA strains. This result might be attributed to antibiotic pressure, facilitating clonal selection. [ABSTRACT FROM AUTHOR]

Abstract Background The prevalence of Staphylococcus aureus varies depending on the healthcare facility, region and country. To understand its genetic diversity, transmission, dissemination, epidemiology and evolution in a particular geographical location, it is important to understand the similarities and variations in the population being studied. This can be achieved by using various molecular characterisation techniques. This study aimed to provide detailed molecular characterisation of South African mecA-positive S. aureus blood culture isolates by describing the SCCmec types, spa types and to lesser extent, the sequence types obtained from two consecutive national surveillance studies. Methods S. aureus blood culture isolates from a national laboratory-based and enhanced surveillance programme were identified and antimicrobial susceptibility testing was performed using automated systems. A real-time PCR assay confirmed the presence of the methicillin-resistance determinant, mecA. Conventional PCR assays were used to identify the SCCmec type and spa type, which was subsequently analysed using the Ridom StaphType™ software. Multilocus sequence typing was performed on selected isolates using conventional methods. MRSA clones were defined by their sequence type (ST), SCCmec type and spa type. Results A detailed description of findings is reported in this manuscript. SCCmec type III predominated overall followed by type IV. A total of 71 different spa types and 24 novel spa types were observed. Spa type t037 was the most common and predominated throughout followed by t1257. Isolates were multidrug resistant; isolates belonging to all SCCmec types were resistant to most of the antibiotics with the exception of type I; isolates with spa type t045 showed resistance to all antibiotics except vancomycin. The most diverse SCCmec-spa type complex was composed of the SCCmec type IV element and 53 different spa types. Conclusion Although ST data was limited, thereby limiting the number of clones that could be identified, the circulating clones were relatively diverse.

Abstract Background The global spread of methicillin-resistant Staphylococcus aureus (MRSA) infections necessitates the use of validated methods for the identification and typing of this bacterium. This study aimed to determine the distribution of main molecular types of MRSA strain circulating among hospitalized patients in teaching hospitals in Isfahan and Kashan. Methods A total of 146 Staphylococcus aureus strains were isolated from patients in four teaching hospitals in Isfahan and Kashan during June 2017 to September 2018. The antimicrobial resistance patterns of Staphylococcus aureus strains were performed by disc diffusion method. The MRSA strains were identified phenotypically and confirmed by PCR assay. The prevalence of microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) genes among MRSA strains was evaluated by multiplex PCR. The genotypes of MRSA strains were determined by multilocus sequence typing and SCCmec typing. Results Of 146 Staphylococcus aureus isolates, 24 (16.4%) isolates were identified as MRSA strains. According to antimicrobial susceptibility testing the highest resistance rates were seen for tetracycline, erythromycin, ciprofloxacin and gentamicin. All of Staphylococcus aureus isolates were susceptible to vancomycin whereas 3 (2.1%) isolates were resistant to linezolid. Three different SCCmec types were obtained among MRSA strains including 16 (66.7%) SCCmec type V, 3 (12.5%) SCCmec type III and 5 (20.8%) SCCmec type II. Of 24 MRSA isolates 20 (83.3%) carried MSCRAMMs genes including eno (70.8%), fib (54.1%), cna (25.0%), fnbB (16.6%), ebps 5 (20.8%), and the fnbA, bbp and clfA genes were not detected in any MRSA isolate. MLST analysis revealed 11 sequence types among MRSA isolates as follows: ST239, ST291, ST22, ST861, ST889, ST8, ST59, ST343, ST772, ST6 and ST1465. Also seven MLST-based clonal complexes (CCs) were identified among MRSA strains including: CC8, CC7, CC398, CC59, CC22, CC1 and CC5. Conclusions A relatively high diversity was found in MRSA genotypes in Kashan and Isfahan hospitals, and seven clonal complexes were identified. Pandemic MRSA clones including CC8 and CC22 were the most prevalent clones and the novel ST types including ST1465, ST861, ST 889 and ST772 are reported for the first time in Iran in the present study. In addition the high prevalence of MSCRAMMs genes in MRSA isolates demonstrates the high potential of these strains for pathogenicity.

Roghayeh Samadi,1 Zohreh Ghalavand,1 Reza Mirnejad,2 Bahram Nikmanesh,3 Gita Eslami1 1Department of Microbiology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran; 2Molecular Biology Research Center, Systems Biology and Poisoning Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran; 3Department of Medical Laboratory Sciences, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, IranCorrespondence: Gita EslamiDepartment of Microbiology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, IranTel +98-2123872556Email g_eslami@yahoo.comIntroduction: Methicillin-resistant Staphylococcus aureus (MRSA) causes high rates of mortality and a substantial burden to health systems worldwide. Here, we investigated the antimicrobial susceptibility and molecular characteristics of MRSA isolated from children referred to Children’s Medical Center in Tehran.Materials and methods: A total of 98 MRSA isolates were collected from children. Antimicrobial resistance patterns were determined using the disk diffusion and E-test methods. The presence of biofilm encoding genes and the pvl gene were determined by PCR. We used the microtiter plate method to assess the ability of biofilm formation. The MRSA isolates were further analyzed using PFGE and SCCmec typing.Results: Antibiotic susceptibility testing showed that the highest and the lowest antibiotic resistance percentage were related to erythromycin (62%) and minocycline (10%), respectively. Overall, 63% of MRSA isolates were biofilm producers. Resistance to two antibiotics such as erythromycin (72% vs 28%, P=0.01) and clindamycin (71% vs 29%, P=0.04) was higher among biofilm producers than non-biofilm producers. All strains had biofilm-forming genes and the prevalence of pvl gene was 41%. Most MRSA isolates belonged to SCCmec IVa (75%) and SCCmec III (18%). In PFGE technique, 5 common types and 2 single types were identified; Common type 1 with 37 isolates was dominant clone.Conclusion: We thus report preliminary data on the prevalence and distribution of MRSA genotypes in Tehran Children’s Hospital. These findings characterize the MRSA colonization dynamics in child patients in Iran and may aid the design of strategies to prevent MRSA infection and dissemination.Keywords: MRSA, biofilm formation, antibiotic resistance, SCCmec typing, PFGE

Background & Objective: Diabetic foot ulcer (DFU), is one of the most frequent causes for hospitalizations in patients with diabetes. A major problem in the treatment of DFU is the increased-incidence of methicillin-resistant Staphylococcus aureus (MRSA). The aim of this study was to determine the SCCmec types of MRSA isolates and their epidemiology among patients with diabetes. Methods: This study was carried out on 145 diabetic patients with DFUs. The antibiotic susceptibility tests (ASTs) were performed using the disk diffusion method and E-test technique. SCCmec typing was done by multiplex PCR. Moreover, the presence of virulence toxin genes, including pvl and lukED was detected by PCR assay. Result: In 145 samples from which S. aureus was predominantly isolated, 19.48% were MRSA. Analysis of MRSA isolates revealed that the most prevalent SCCmec type was type IV (46.7%) followed by type III (30.0%) and type V (20.0%). One strain (3.3%) was untypeable. The prevalence of pvl and lukED was 56.7% and 100%, respectively. Conclusion: The high prevalence of MRSA in DFUs represents the high levels of antibiotic usage among patients with diabetes. In this study, resistance to other important clinical antibiotics was detected among MRSA isolates. The high proportion of SCCmec type IV and V strains, even in former hospitalized patients, indicates the entrance of these clones to the clinical setting.

Background and purpose: The aims of this study were to evaluate the antibiotic resistance of Healthcare-associated Methicillin-resistant Staphylococcus aureus (HA-MRSA) and Community associated MRSA (CA-MRSA)-resistant strains of Staphylococcus aureus strains and to investigate the frequency of different types of spa typing, SCCmec I, II, III, IV, V, and type IV, among the strains of MRSA isolated from clinical specimens of children in Tehran Children's Medical Center Hospital. Materials and methods: In this cross-sectional study, antibiotic susceptibility test was performed using Kirby-Baer method according to the CLSI guidelines. Presence of mecA, PVL, spa genes, as well as different types of SCCmec I, II, III, IV, V, and major subtypes of SCCmec IV were investigated using Multiplex PCR. Results: Among 133 clinical isolates of S. aureus, 70 (53%) were methicillin-susceptible (MSSA) and 63 (47%) were MRSA. The mecA and PVL genes frequency distributions among MRSA strains were 100% and 3.1%, respectively. In the present study, there were 5 SCCmec (I-V) types, indicating wide variations in the hospital investigated. In addition, high prevalence of SCCmec III (30.1%) and III + Iva (23.8%) was observed. Of 63 strains of MRSA, 46 were classified into 11 spa types. The most common type was spa t037 that was found in 53.9% (n= 34) of the strains. Conclusion: High prevalence of SCCmec III + Iva and emergence of CA-MRSA strains in hospitals can be a serious warning to practitioners and infection control committees.

Abstract Objectives Molecular typing such as spa typing is used to control and prevent Staphylococcus aureus widespread in hospitals and communities. Hence, the aim of this study was to find the most common types of S. aureus strain circulating in Shiraz via spa and SCCmec typing methods. Results Total of 159 S. aureus isolates were collected from two tertiary hospitals in Shiraz. Isolates were identified by biochemical tests. Antimicrobial susceptibility tests were performed by standard disk diffusion method and then genetic analysis of bacteria was performed using SCCmec and spa typing. In this study 31.4% of the isolates were methicillin-resistant S. aureus. The majority of isolates were SSCmec type III. Spa type t030 was the most prominent type among MRSA strains. For the first time in Iran, spa003, t386, t1877, t314, t186, t1816, t304, t325, t345 were reported in this study. It was shown that there is a possibility that these spa types are native to this region. Our findings showed that SCCmec II, III and IV disseminate from hospital to community and vice versa. Thus, effective monitoring of MRSA in hospital and community is necessary.

Background: SCCmec and PCR-RFLP are productive and cost-effective methods for epidemiological investigations and source tracking of MRSA. Aim of this study was to investigate the epidemiology and molecular characterization of MRSA recovered from device related infection. Methods: A total of 626 MRSA were collected from prosthetic device related infections and subjected for presence of mec gene and SCCmec typing. SCCmec characterized MRSA were subjected for agr typing followed by RFLP genotyping and amplification of PVL gene. Following the Clinical & Laboratory Standards Institute (CLSI), antibiotic resistance pattern was determined. Results: Of 626 MRSA isolates, 488(78%) were characterized by SCCmec typing. Most common type was SCCmec IV (43%), followed by SCCmec II (25%) SCCmec III (22%) and only 10% was SCCmec V. SCCmec characterized MRSA strains were 100% resistance to tobramycin and chloramphenicol, 96% resistance to ciprofloxacin and 93% resistance to azithromycin. The gene PVL was only present in SCCmec IV. All agr typed MRSA strains were resistance to gentamicin, tobramycin and chloramphenicol. PVL was present in all strains of agr III, 80% of agrIV, 50% of agrII and absent in agrI. RFLP analysis resulted in 16 types of non-duplicate unique bands pattern which were equally distributed among prosthetic device related infection. Conclusion: PVL harboring SCCmec or agr typed MRSA strains are less resistance to antibiotics. RFLP is simple, productive and cost-effective method for molecular typing of MRSA. Correct use of antibiotics and molecular surveillance is indispensable to detect the change in epidemiological and antibiotic resistance trends of MRSA.

Widespread introduction of HIV integrase inhibitors into clinical care may result in appearance of drug resistance mutations affecting treatment outcome. The aim of our study was to monitor the resistance patterns of integrase inhibitors beside protease and reverse transcriptase inhibitors in newly diagnosed therapy-naive HIV-positive patients in Hungary between 2017 and 2019. Genotype-based resistance testing of HIV integrase, protease and reverse transcriptase was performed by amplification and Sanger population sequencing from plasma samples. Drug resistance mutations were identified by the algorithm of Stanford HIV Drug Resistance Database. Potentially transmitted, non-polymorphic integrase major mutation was detected in 1 out of 249 samples, while accessory mutations were observed in further 31 patients (12.4%). The overall prevalence of transmitted drug resistance (TDR) mutations related to protease and reverse transcriptase inhibitors was 5.8% (10/173) between the end of 2017 and 2019. Nucleoside reverse transcriptase inhibitor associated resistance mutations were the most frequent indicators of TDR (6/173; 3.5%), followed by resistance mutations associated with protease (3/173; 1.7%) and non-nucleoside reverse transcriptase inhibitors (2/173, 1.2%). The first detection of integrase major mutation and the changing patterns of other resistance mutations in Hungarian untreated HIV-positive population indicate the necessity of continuous molecular surveillance of Hungarian HIV epidemic. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is a common pathogen causing hospital infections. The increasing rate of healthcare-associated infections caused by methicillin-resistant Staphylococcus aureus (MRSA) in developing countries has led to many public health problems. This study aimed to investigate the molecular epidemiology as well as the antibiotic resistance pattern of clinical isolates of MRSA from Southern Iran. A total of 135 S. aureus isolates were collected from the patients referred to three hospitals in South Iran. The phenotypic and genotypic diagnosis of MRSA isolates was performed by disk diffusion and PCR methods, respectively. The antibiotic resistance pattern for MRSA isolates was performed using Kirby–Bauer method. The molecular epidemiology of isolates was performed by MLST, Spa typing and SCC mec typing. From 135 S. aureus isolates, 50 (37%) MRSA strains were detected from which two different sequence types including ST239 and ST605 were identified. SCC mec type III was the most common profile (50%) and t030 was the predominant spa type (48%) among the strains. The MRSA isolates had the highest resistance to penicillin (100%), tetracycline (88%), levofloxacin (86%), ciprofloxacin (84%), erythromycin (82%), gentamicin (80%), and clindamycin (78%). The results of this study show that the most common genetic type among the MRSA isolates was ST239-SCC mec III/t030. The rapid and timely detection of MRSA and the administration of appropriate antibiotics according to the published antibiotic resistance patterns are essential. Furthermore, the continuous and nationwide MRSA surveillance studies are necessary to investigate clonal distribution and spreading of MRSA from community to hospitals. [ABSTRACT FROM AUTHOR]

Multidrug-resistant Staphylococcus aureus strains have been commonly found in hospitals and communities causing wide ranges of infections among humans and animals. Typing of these strains is a key factor to reveal their clonal dissemination in different regions. We investigated the prevalence and dissemination of different clonal groups of S. aureus with resistance phenotype to multiple antibiotics in two sewage treatment plants (STPs) in Tehran, Iran over four sampling occasions. A total of 576 S. aureus were isolated from the inlet, sludge and outlet. Of these, 80 were identified as methicillin-resistant S. aureus (MRSA) and were further characterized using a combination of Phene Plate (PhP) typing, staphylococcal cassette chromosome mec (SCCmec), ccr types, prophage and antibiotic-resistant profiling. In all, eight common type (CT) and 13 single PhP type were identified in both STPs, with one major CT accounting for 38.8% of the MRSA strains. These strains belonged to three prophage patterns and five prophage types with SCCmec type III being the predominant type. Resistance to 11 out of the 17 antibiotics tested was significantly (P< 0.0059) higher among the MRSA isolates than methicillin-sensitive S. aureus (MSSA) strains. The persistence of the strains in samples collected from the outlet of both STPs was 31.9% for MRSA and 23.1% for MSSA. These data indicated that while the sewage treatment process, in general, is still useful for removing most MRSA populations, some strains with SCCmec type III may have a better ability to survive the STP process. [ABSTRACT FROM AUTHOR]

Background Coagulase-negative staphylococci (CoNS) are emerging as clinically significant pathogens. A high proportion of methicillin resistance along with intense biofilm-producing ability render CoNS-related infections challenging to treat. This study was undertaken to investigate the mechanisms of methicillin resistance, identify genes encoding for virulence, and their association with clinical outcomes among clinical isolates of Staphylococci in a tertiary care center. Methods A total of 203 clinical isolates were included in this study. Susceptibility to various antibiotics was determined by the disc diffusion method. Methicillin resistance was screened using cefoxitin disc, mecA  and mecC genes were detected using polymerase chain reaction (PCR). PCR was performed to detect five virulence genes:  atlE , aap , fbe , embp, and icaAB . Staphylococcal cassette chromosome mec (SCCmec) types were identified by multiplex PCR. Statistical analysis was performed using SPSS software (IBM Inc., Armonk, New York). The Chi-squared test was used to compare the distribution of virulence genes among methicillin-susceptible resistant CoNS. A p-value of less than 0.5 was considered significant. Results In the current study, 60% (122/203) of CoNS were methicillin-resistant, and SCCmec type I was the most common. Among the 203 CoNS, 24.6% (50/203) isolates harbored one or more virulence genes in them.  Conclusion CoNS have relatively low virulence as only 24.6% of isolates carried the virulence genes. Nevertheless, the variety of diseases linked to these species indicates the necessity for accurate identification and precise reporting of antimicrobial susceptibility to avoid adverse outcomes., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. Institutional Ethics Committee of Sri Ramachandra Institute of Higher Education and Research issued approval IEC-NI/19/FEB/68/12. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Khodabux et al.)

Abstract Background The emergence of methicillin-resistant Staphylococcus aureus (MRSA) is a main concern in burn care centers worldwide. The some reports of MRSA in Iran suggested that MRSA with type SCCmec III is common among burn patients. The aim of this study was to determine the prevalence, virulence genes, and antimicrobial susceptibility of the direct repeat units (dru) types of MRSA with SCCmec IIIA isolated from burn wounds in a burn care center in Tehran, Iran. Methods In total, 165 S. aureus isolates were collected from clinical samples. In order to detect MRSA isolates, the mecA gene was amplified through the polymerase chain reaction (PCR) method. Antimicrobial susceptibility was tested using the disc agar diffusion test. Moreover, the PCR method was applied to determine SCCmec types, virulence genes, and antimicrobial resistance genes. The dru region was sequenced and thereby, dru types and dru repeats were identified. A similarity matrix was used to create minimum spanning tree (MST). Results The prevalence of MRSA was 69% (114 out of 165 isolates). Most of MRSA isolates (61 out of 114, 53.5%) were SCCmec type IIIA. All MRSA isolates were vancomycin-susceptible and more than 68% of MRSA isolates with SCCmec type IIIA were mupirocin resistant. The successful dru typing of isolates with SCCmec type IIIA revealed fourteen different dru types. There were two new dru types, namely dt10di and dt7aj. MST analysis indicated the presence of the three clusters of dt10di (cluster I), dt8i-dt8 h (cluster II), and dt11c-dt10ao-dt11dd-dt11a-dt10a (cluster III). There were significant differences between clusters I and II respecting antimicrobial resistance pattern and virulence genes. Conclusion Three main dru clusters are prevalent in the study setting. The main dru types in the setting are dt10di, dt8i, and dt8 h. Dru typing can be used to differentiate MRSA strains with SCCmec IIIA.

Background: The global spread of methicillin-resistant Staphylococcus aureus (MRSA) infections necessitates the use of validated methods for the identification and typing of this bacterium. This study aimed to determine the distribution of main molecular types of MRSA strain circulating among hospitalized patients in teaching hospitals in Isfahan and Kashan. Methods: A total of 146 Staphylococcus aureus strains were isolated from patients in four teaching hospitals in Isfahan and Kashan during June 2017 to September 2018. The antimicrobial resistance patterns of Staphylococcus aureus strains were performed by disc diffusion method. The MRSA strains were identified phenotypically and confirmed by PCR assay. The prevalence of microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) genes among MRSA strains was evaluated by multiplex PCR. The genotypes of MRSA strains were determined by multilocus sequence typing and SCCmec typing. Results: Of 146 Staphylococcus aureus isolates, 24 (16.4%) isolates were identified as MRSA strains. According to antimicrobial susceptibility testing the highest resistance rates were seen for tetracycline, erythromycin, ciprofloxacin and gentamicin. All of Staphylococcus aureus isolates were susceptible to vancomycin whereas 3 (2.1%) isolates were resistant to linezolid. Three different SCCmec types were obtained among MRSA strains including 16 (66.7%) SCCmec type V, 3 (12.5%) SCCmec type III and 5 (20.8%) SCCmec type II. Of 24 MRSA isolates 20 (83.3%) carried MSCRAMMs genes including eno (70.8%), fib (54.1%), cna (25.0%), fnbB (16.6%), ebps 5 (20.8%), and the fnbA, bbp and clfA genes were not detected in any MRSA isolate. MLST analysis revealed 11 sequence types among MRSA isolates as follows: ST239, ST291, ST22, ST861, ST889, ST8, ST59, ST343, ST772, ST6 and ST1465. Also seven MLST-based clonal complexes (CCs) were identified among MRSA strains including: CC8, CC7, CC398, CC59, CC22, CC1 and CC5. Conclusions: A relatively high diversity was found in MRSA genotypes in Kashan and Isfahan hospitals, and seven clonal complexes were identified. Pandemic MRSA clones including CC8 and CC22 were the most prevalent clones and the novel ST types including ST1465, ST861, ST 889 and ST772 are reported for the first time in Iran in the present study. In addition the high prevalence of MSCRAMMs genes in MRSA isolates demonstrates the high potential of these strains for pathogenicity. [ABSTRACT FROM AUTHOR]

Staphylococcal cassette chromosome mec (SCCmec) typing was established in the 2000s and has been employed as a tool for the molecular epidemiology of methicillin-resistant Staphylococcus aureus, as well as the evolution investigation of Staphylococcus species. Molecular cloning and the conventional sequencing of SCCmec have been adopted to verify the presence and structure of a novel SCCmec type, while convenient PCR-based SCCmec identification methods have been used in practical settings for many years. In addition, whole-genome sequencing has been widely used, and various SCCmec and similar structures have been recently identified in various species. The current status of the SCCmec types, SCCmec subtypes, rules for nomenclature, and multiple methods for identifying SCCmec types and subtypes were summarized in this review, according to the perspective of the International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements.

Background: Asymptomatic nasal colonization of Methicillin-Resistant Staphylococcus aureus is common in Multiple Sclerosis patients. SCCmec types I to III are mainly attributed to HA-MRSA strains whereas SCCmec types IV and V have commonly been reported in CA-MRSA infections. Here, we assessed the frequency of nasal carriage of MRSA in MS patients. This study aimed to evaluate MRSA SCCmec typing in MS nasal carriage. Methods: A cross-sectional descriptive study was conducted from Feb and Jun 2017 in MS Research Center, Tehran University of Medical Sciences (TUMS), Iran. Overall, 620 nasal swabs were collected (325 from MS patients and 295 from control group). Antimicrobial susceptibility test was performed using the disk diffusion and E-test method. Presence of mecA gene was confirmed by PCR assay and multiplex PCR was performed for SCCmec typing of MRSA isolates. Results: The frequency of MRSA among the MS patients and control group was almost equal (9.2% and 10.1%, respectively). SCCmec typing detected only types III, IV and V in both groups and type IV was the most predominant type in MS patients and control group. SCCmec type III was more prevalent in control group than MS patients (40% vs. 20%). Moreover, the frequency of SCCmec type V in MS patients was significantly higher than control group (36.7% vs. 3.3%). Conclusion: Although most MRSA isolates were collected from inpatients, interestingly there is a high frequency of SCCmec types IV and V in MS group. Moreover, MRSA isolates were not resistant to more antibiotics in SCCmec type III than types IV-V.

Resistance to methicillin in methicillin resistant Staphylococcus aureus (MRSA) is dependent on mecA gene located on staphylococcal cassette chromosome (SCC). Both SCCmec type and Panton-Valentine leukocidin (PVL) affect S. aureus pathogenicity. Aim of this study was to investigate the prevalence of SCCmecA types and pvl genes among MRSA isolates from inpatients. During this cross-sectional study on 100 clinical isolates, following antibiotic susceptibility test, screening of mecA and pvl genes, as well as SCCmec typing, was done in a multiplex PCR technique. From the studied samples, 58 isolates were recognized as MRSA. The frequency of mecA and pvl was 58% and 4%, respectively. All of the MRSA were resistant to cefoxitin and had the highest sensitivity to chloramphenicol. The majority (77.5%) of MRSA was originated from wound samples. The SCCmec III was the most frequent type (22.4%) in these samples. The pvl positive isolates were from SCCmec IVb and V, thus meaning they are from CA-MRSA. These results show a high prevalence of MRSA in the studied region and a widespread prevalence of SCCmec I-V types. Furthermore, high prevalence of SCCmec III indicates the prevalence of multidrug resistant MRSA. This finding is a serious alarm for medical health care practitioners for the correct use of antibiotics in order to limit the spread of multidrug resistant strains. In addition, with regard to life threatening infections caused by pvl harbouring strains, early diagnosis and treatment of infections caused by these isolates should be mandatory.

• MRSA nasal carriage of Syrian refugees were higher than that found in Western countries; final destination of refugees. • The most common SCC mec type in Syrian refugees and host community host and refugees was types IVa. • SCC mec types of the host community were different from previous results in the region • Such changes in genetic makeup may highlight the possibility of substantial evolution in MRSA within a short period. The aim was to investigate methicillin-resistant Staphylococcus aureus (MRSA) carriage rates and to characterize the staphylococcal cassette chromosome mec (SCC mec) among Syrian refugees and the host community in Duhok, Iraq. A total of 492 host community and 355 Syrian refugee subjects were recruited. Participant bio-information was collected using a study questionnaire. MRSA carriage was identified as recommended in the Clinical and Laboratory Standards Institute guidelines. PCR was performed for typing the mec A gene and SCC mec groups. In the host community, 76/492 (15.4%) carried MRSA, whereas 49/355 (13.8%) Syrian refugees carried MRSA (p = 0.505). Refugees were classified according to their arrival date; 36/278 (13%) existing refugees and 13/77 (16.9%) new refugee arrivals were identified as MRSA carriers (p = 0.375). Regarding the risk factors influencing MRSA spread, no association was found between MRSA prevalence and risk factors. The most common types of SCC mec in both communities were types IVa and V: 67.1% and 5.26%, and 49% and 2.04%, respectively. It is inferred that the effect of MRSA in refugees on the host community will be negligible. However, the refugee carriage rate is higher than that found in Western countries, the final destination of refugees. Therefore, screening for MRSA is mandated and the rapid detection of carriers would prevent its spread. [ABSTRACT FROM AUTHOR]

Purpose: This in-vitro study aimed to assess in 120 [40 community-acquired (CA-MRSA) & 80 hospital-acquired (HA-MRSA)] isolates from cancer patients whether the transmissible staphylococcal cassette chromosome mec (SCCmec) typing, and the Panton–Valentine leukocidin (PVL) virulence genes detection could be employed as tools for molecular diagnostic purposes to distinguish both methicillin-resistant Staphylococcus aureus (MRSA) categories in radiotherapy treated cancer patients. Materials and methods: SCCmec typing was determined by the combination of the type of the cassette chromosome recombinase genes (ccr) gene complex and the class of the methicillin resistance (mec) gene complex. Besides, a rapid slide latex agglutination test (LAT) and antibiotic resistance spectrum determination before and after irradiation were performed. Results: In the strict sense, with the effect of irradiation; the presence of SCCmec subtypes IVa (22.5% vs. 10.0%), b (47.5% vs. 25.0%), & d (7.5 vs. 2.5%) or type V (15.0% vs. 7.5%) genetic elements and PVL genes (p <.001) were not proved as a signature for CA-MRSA. While, the larger SCCmec types II, and III elements were not detected in 14, and 19 from the 38, and 36 typed HA-MRSA isolates (p <.001), respectively. Remarkable effects on class A & class B mec gene complex and type2, type 3 & type 5 ccr gene complex and an increase in agglutination reaction strength in response to gamma irradiation external stimulus were observed. Conclusions: Different heterogeneous genetic composition with upregulation mecA gene expression was detected after irradiation in the HA- MRSA studied population. CA-MRSA showed remarkable ability to acquire multi-antibiotic resistance after irradiation and propose a novel paradigm for future chemotherapy against the multi-resistant pathogens whose proliferation especially among immunocompromised cancer patients is on the increase. [ABSTRACT FROM AUTHOR]

Background & Objective: Diabetic foot ulcer (DFU), is one of the most frequent causes for hospitalizations in patients with diabetes. A major problem in the treatment of DFU is the increased-incidence of methicillin-resistant Staphylococcus aureus (MRSA). The aim of this study was to determine the SCCmec types of MRSA isolates and their epidemiology among patients with diabetes. Methods: This study was carried out on 145 diabetic patients with DFUs. The antibiotic susceptibility tests (ASTs) were performed using the disk diffusion method and E-test technique. SCCmec typing was done by multiplex PCR. Moreover, the presence of virulence toxin genes, including pvl and lukED was detected by PCR assay. Results: In 145 samples from which S. aureus was predominantly isolated, 19.48% were MRSA. Analysis of MRSA isolates revealed that the most prevalent SCCmec type was type IV (46.7%) followed by type III (30.0%) and type V (20.0%). One strain (3.3%) was untypeable. The prevalence of pvl and lukED was 56.7% and 100%, respectively. Conclusion: The high prevalence of MRSA in DFUs represents the high levels of antibiotic usage among patients with diabetes. In this study, resistance to other important clinical antibiotics was detected among MRSA isolates. The high proportion of SCCmec type IV and V strains, even in former hospitalized patients, indicates the entrance of these clones to the clinical setting. [ABSTRACT FROM AUTHOR]

Resistance to methicillin in methicillin resistant Staphylococcus aureus (MRSA) is dependent on mecA gene located on staphylococcal cassette chromosome (SCC). Both SCCmec type and Panton-Valentine leukocidin (PVL) affect S. aureus pathogenicity. Aim of this study was to investigate the prevalence of SCCmecA types and pvl genes among MRSA isolates from inpatients. During this cross-sectional study on 100 clinical isolates, following antibiotic susceptibility test, screening of mecA and pvl genes, as well as SCCmec typing, was done in a multiplex PCR technique. From the studied samples, 58 isolates were recognized as MRSA. The frequency of mecA and pvl was 58% and 4%, respectively. All of the MRSA were resistant to cefoxitin and had the highest sensitivity to chloramphenicol. The majority (77.5%) of MRSA was originated from wound samples. The SCCmec III was the most frequent type (22.4%) in these samples. The pvl positive isolates were from SCCmec IVb and V, thus meaning they are from CA-MRSA. These results show a high prevalence of MRSA in the studied region and a widespread prevalence of SCCmec I-V types. Furthermore, high prevalence of SCCmec III indicates the prevalence of multidrug resistant MRSA. This finding is a serious alarm for medical health care practitioners for the correct use of antibiotics in order to limit the spread of multidrug resistant strains. In addition, with regard to life threatening infections caused by pvl harbouring strains, early diagnosis and treatment of infections caused by these isolates should be mandatory. [ABSTRACT FROM AUTHOR]

To provide evidence of the cross-contamination of emerging pathogenic microbes in a local network between long-term care facilities (LTCFs) and hospitals, this study emphasizes the molecular typing, the prevalence of virulence genes, and the antibiotic resistance pattern of methicillin-resistant Staphylococcus aureus. MRSA isolates were characterized from 246 samples collected from LTCFs, medical tubes of LTCF residents, and hospital environments of two cities, Chiayi and Changhua. Species identification, molecular characterization, and drug resistance analysis were performed. Hospital environments had a higher MRSA detection rate than that of LTCF environments, where moist samples are a hotspot of MRSA habitats, including tube samples from LTCF residents. All MRSA isolates in this study carried the exfoliative toxin eta gene (100%). The majority of MRSA isolates were resistant to erythromycin (76.7%), gentamicin (60%), and ciprofloxacin (55%). The percentage of multidrug-resistant MRSA isolates was approximately 50%. The enterobacterial repetitive intergenic consensus polymerase chain reaction results showed that 18 MRSA isolates belonged to a specific cluster. This implied that genetically similar isolates were spread between hospitals and LTCFs in Changhua city. This study highlights the threat to the health of LTCFs’ residents posed by hospital contact with MRSA.

Staphylococcus aureus is one of the major causes of bloodstream infections. The aim of our study was to characterize methicillin-resistant Staphylococcus aureus (MRSA) isolates from blood of patients hospitalized in the Czech Republic between 2016 and 2018. All MRSA strains were tested for antibiotic susceptibility, analyzed by spa typing and clustered using a Based Upon Repeat Pattern (BURP) algorithm. The representative isolates of the four most common spa types and representative isolates of all spa clonal complexes were further typed by multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) typing. The majority of MRSA strains were resistant to ciprofloxacin (94%), erythromycin (95.5%) and clindamycin (95.6%). Among the 618 strains analyzed, 52 different spa types were detected. BURP analysis divided them into six different clusters. The most common spa types were t003, t586, t014 and t002, all belonging to the CC5 (clonal complex). CC5 was the most abundant MLST CC of our study, comprising of 91.7% (n = 565) of spa-typeable isolates. Other CCs present in our study were CC398, CC22, CC8, CC45 and CC97. To our knowledge, this is the biggest nationwide study aimed at typing MRSA blood isolates from the Czech Republic.

This study characterized EMRSA-15 isolates obtained from patients in Kuwait hospitals for their genotypic relatedness, antibiotic resistance and carriage of virulence genes using pulsed-field gel electrophoresis (PFGE), coagulase serotyping, SCCmec subtyping, spa typing, multilocus sequence typing and DNA microarray. The isolates were resistant to trimethoprim (75.6%), ciprofloxacin (29.7%), erythromycin and clindamycin (24.3%), tetracycline (19.0%), and gentamicin and kanamycin (21.6%). All 37 isolates belonged to sequence type (ST) 22, coagulase type XI, three PFGE types and eight subtypes, ten spa types including t223 (51.3%), t852 (13.5%), t032 (8.1%), t790 (8.1%), t3107 (5.4%) and one each of t309, t2251, t3935, t5708 and t5983. Twenty-six isolates (70.2%) carried SCCmec IVa, eight isolates carried SCCmec IV and three isolates carried SCCmec IVh. All isolates carried agr1, cap5 and egc gene cluster (seg, sei, selm, seln, selo, and selu). tst (toxic shock syndrome toxin) was detected in 23 isolates. Eight isolates (21.6%) were positive for Panton-Valentine leukocidin (PVL). Genotypic analysis revealed that 62.1% of the isolates comprising ST22-IVa-t223 (51.3%) and ST22-IVa-t309/t2251/t3935/t5708 (10.8%) were CC22-[tst1+] UK EMRSA-15/Middle Eastern variant, 21.6% were CC22-PVL+ EMRSA-15 variant and 16.2% were CC22-UK EMRSA-15/Barnim clone. These results show that the tst1 positive-ST22-IVa-t223 (Middle Eastern variant) and the CC22-PVL+ EMRSA-15 variant were the dominant EMRSA-15 variants in Kuwait hospitals.

Periodical surveillance on nosocomial pathogens is important for antimicrobial stewardship and infection control. The first methicillin-resistant Staphylococcus aureus (MRSA) molecular surveillance in Hospital Canselor Tuanku Muhriz (HCTM), a Malaysian teaching hospital, was performed in 2009. The dominant clone was identified as an MRSA carrying SCCmec type III-SCCmercury with ccrC and sea+cna toxin genes. In this study, we report the findings of the second HCTM MRSA surveillance carried out in 2017, after an interval of 8 years. Antibiotic susceptibility testing, SCCmec, toxin gene, and spa typing were performed for 222 MRSA strains isolated in 2017. Most strains were resistant to ciprofloxacin, erythromycin, clindamycin, cefoxitin, and penicillin (n = 126, 56.8%), belong to SCCmec type IV (n = 205, 92.3%), spa type t032 (n = 160, 72.1%) and harboured seg+sei toxin genes (n = 172, 77.5%). There was significant association between resistance of the aforementioned antibiotics with SCCmec type IV (p < 0.05), t032 (p < 0.001), and seg+sei carriage (p < 0.05). Results from this second MRSA surveillance revealed the occurrence of clonal replacement in HCTM during an interval of not more than 8 years. Investigation of the corresponding phenotype changes in this new dominant MRSA clone is currently on-going.

The community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has become increasingly prevalent in both community and hospital settings. The aim of this study was to determine the prevalence, molecular characteristics and antibiotic resistance profiles of CA-MRSA from community- and hospital-associated infections in a tertiary care hospital in Mangalore, India. Of 520 S. aureus isolates, 362 were from inpatients (IP) and 158 were from outpatients (OP). One-hundred and thirty-two MRSA isolates obtained from 94 inpatients and 38 outpatients with complete clinical details were further analyzed. Of these, 81 (61.4%) were CA-MRSA (IP-47.9%, OP-94.7%) and 51 (38.6%) were HA-MRSA (IP-52.1%, OP-5.3%). All (100%) MRSA isolates were mecA gene positive. SCCmec typing identified SCCmec type IV (50.6%) and SCCmec type V (66.7%) in CA-MRSA, while SCCmec type I (41.2%), SCCmec type III (19.6%), SCCmec type IV (31.4%) and SCCmec type V (25.5%) were detected in HA-MRSA isolates. The Panton–Valentine Leukocidin (PVL) gene was found in 70.4% of CA-MRSA, 43.1% of HA-MRSA with SCCmec type IV and SCCmec type V, and in 7.8% of true HA-MRSA. The antibiotic resistance profiles were determined by the disc diffusion method. Resistance to cefoxitin was used to identify MRSA. A significant difference (p < 0.05) was observed between CA-MRSA and HA-MRSA with respect to resistance against cephalexin, cefotaxime, levofloxacin, linezolid and teicoplanin. CA-MRSA was predominantly resistant to ciprofloxacin (86.4%), erythromycin (66.7%), ofloxacin (49.4%), cefotaxime (44.4%), gentamicin (40.7%) and clindamycin (40.7%), while HA-MRSA showed resistance against ciprofloxacin (80.4%), erythromycin (80.1%), cefotaxime (70.6%),ofloxacin (58.8%), clindamycin (47.1%) and levofloxacin (41.2%).This study reports the prevalence of CA-MRSA in community and hospital settings and the possibility of multidrug-resistant CA-MRSA replacing HA-MRSA in hospitals. The observations from our study emphasize the need for urgent measures to manage this emerging crisis in healthcare settings.

Background and Objectives: Staphylococcus aureus is an important pathogen that can be colonized in the nose and increase the risk of spreading infections in hospitals. The present study aimed at determining the phenotypic and genotypic characterization of S. aureus strains isolated from patients and healthcare workers (HCWs) from a teaching hospital in Isfahan, Iran. Materials and Methods: This cross-sectional study was performed on 262 nasal swabs and 23 clinical isolates that were collected from a teaching hospital during February and April 2016. Staphylococcal cassette chromosome mec (SCCmec) and multilocus sequence typing (MLST) were performed for selected isolates. Results: Overall, 23% and 18% of healthcare workers and patients were carriers, respectively. Methicillin-resistant S. aureus (MRSA) rate was 13%, 33% and 52% in nasal HCWs, nasal patients, and clinical samples, respectively. The molecular typing of MRSA isolates revealed that the most common SCCmec type is SCCmec type III (88%). The highest rate of resistance was observed against tetracycline and erythromycin, with 48.7%. The most frequently detected toxin genes among S. aureus isolates were hla (99%) and sea (44%), moreover, pvl genes were detected in (40%) of MRSA isolates. The results of MLST showed 7 different sequence types (STs): ST859 (2/9), ST6 (2/9), ST639 (1/9), ST343 (1/9), ST239 (1/9), ST291 (1/9) and ST25 (1/9). Conclusion: Our results revealed that ST clones associated with healthcare-associated MRSA (HA-MRSA) are actively circulating among nasal carriage in our healthcare setting, and thus, effective infection control policies are needed to reduce nasal carriage in healthcare settings.

Highlights • 91 S. aureus isolated from 578 mastitis milk samples of buffaloes of three provinces. • Eight different spa types detected with three predominant ones (t3576, t7311, t937). • Four different sequence types (ST97, ST352, ST291, and ST522) detected by MLST. • Two MRSA was detected belonging to t304-ST291-SCC mec IV and t7311-ST522-SCC mec IV. • A significant difference in geographical distribution of genotypes was observed. Abstract The aims of the present study were to investigate the genetic diversity and methicillin resistance in S. aureus isolates recovered from mastitis-affected buffaloes. Five hundred seventy-eight milk samples were obtained from buffaloes with mastitis in three provinces, Iran. Ninety-one of the 578 tested samples contained S. aureus (15.74%), in two cases were methicillin resistant S. aureus (MRSA). Isolates were typed by spa typing, followed by MLST on some representative isolates and SCC mec typing for MRSA strains. The presence of genes encoding Panton–Valentine leukocidin (PVL) was also tested by PCR. Eight spa types were identified, with t3576 (n = 18), t7311 (n = 18) and t937 (n = 17) were the most common, followed by t304 (n = 11), t7308 (n = 9), t521 (n = 7), t267 (n = 6), and t527 (n = 5). MLST revealed four different sequence types (STs) including ST97 (related to t521 and t527 spa types), ST352 (related to t267), ST291 (related to t304 and t937) and ST522 (related to t7338, t7311 and t3576). Two MRSA were identified as t304-ST291-SCC mec IV and t7311-ST522-SCC mec IV. No PVL-positive S. aureus were found. A significant difference in geographical distribution of genotypes was observed, with some types being prevalent in all studied provinces (P < 0.001). The results demonstrated genetic diversity among the S. aureus strains involved in mastitis in buffaloes. This study also provides evidence of the presence of MRSA belonging to genotypes which have been earlier reported in human infections, emphasizing the need for their epidemiological monitoring. [ABSTRACT FROM AUTHOR]

Abstract Infections caused by Staphylococcus aureus remain a major global healthcare problem. We aimed to find the common lineages of S. aureus strains circulating in a burn hospital in Tehran. A total of 167 isolates of S. aureus obtained from patients, healthcare workers (HCWs) and environment in Shahid Motahari burn hospital were genotyped by using spa, agr and staphylococcal cassette chromosome mec (SCC mec) typing methods. Antimicrobial susceptibility testing was performed by using the disc diffusion method. The frequency of methicillin-resistant S. aureus (MRSA) was 64.7% (n = 108), with distribution frequencies among patient, HCW and surface isolates of 64.2% (n = 79), 50% (n = 7) and 73.3% (n = 22), respectively. SCC mec type III (75%, n = 81) was found to be the most frequent SCC mec type among MRSA isolates, followed by SCC mec type I (20.4%, n = 22) and SCC mec type IV (1.8%, n = 2). The remaining MRSA isolates (2.8%, n = 3) were nontypeable by this method. About 78.4% (n = 131), 10.2% (n = 17) and 4.8% (n = 8) of all isolates were characterized as agr types I, II and III, respectively, and the other isolates (6.6%) were nontypeable. spa types t030 and t037 constituted the first and second most predominant spa types found in 56.4% (n = 57) and 25.6% (n = 26) of isolates, respectively. We also report here a novel spa type, t16471. The most prevalent genotypes of the isolates found among patient, surface and HCW samples were SCC mec type III/t030, t037/ agr type I. Continuous tracking of epidemic isolates and better hospital infection control policies are recommended to efficiently prevent the spread of bacteria to inpatients. [ABSTRACT FROM AUTHOR]

Background and Objectives: Staphylococcus aureus is an important pathogen that can be colonized in the nose and increase the risk of spreading infections in hospitals. The present study aimed at determining the phenotypic and genotypic characterization of S. aureus strains isolated from patients and healthcare workers (HCWs) from a teaching hospital in Isfahan, Iran. Materials and Methods: This cross-sectional study was performed on 262 nasal swabs and 23 clinical isolates that were collected from a teaching hospital during February and April 2016. Staphylococcal cassette chromosome mec (SCCmec) and multilocus sequence typing (MLST) were performed for selected isolates. Results: Overall, 23% and 18% of healthcare workers and patients were carriers, respectively. Methicillin-resistant S. aureus (MRSA) rate was 13%, 33% and 52% in nasal HCWs, nasal patients, and clinical samples, respectively. The molecular typing of MRSA isolates revealed that the most common SCCmec type is SCCmec type III (88%). The highest rate of resistance was observed against tetracycline and erythromycin, with 48.7%. The most frequently detected toxin genes among S. aureus isolates were hla (99%) and sea (44%), moreover, pvl genes were detected in (40%) of MRSA isolates. The results of MLST showed 7 different sequence types (STs): ST859 (2/9), ST6 (2/9), ST639 (1/9), ST343 (1/9), ST239 (1/9), ST291 (1/9) and ST25 (1/9). Conclusion: Our results revealed that ST clones associated with healthcare-associated MRSA (HA-MRSA) are actively circulating among nasal carriage in our healthcare setting, and thus, effective infection control policies are needed to reduce nasal carriage in healthcare settings. [ABSTRACT FROM AUTHOR]