Search

Rapid and accurate diagnosis of tuberculosis (TB) is essential to control the disease. The conventional microbiological tests have limitations and there is an urgent need to devise a simple, rapid and reliable point-of-care (POC) test. The failure of TB diagnostic tests based on antibody detection due to inconsistent and imprecise results has stimulated renewed interest in the development of rapid antigen detection methods. However, the World Health Organization (WHO) has emphasized to continue research for designing new antibody-based detection tests with improved accuracy. Immuno-polymerase chain reaction (I-PCR) combines the simplicity and versatility of enzyme-linked immunosorbent assay (ELISA) with the exponential amplification capacity and sensitivity of PCR thus leading to several-fold increase in sensitivity in comparison to analogous ELISA. In this review, we have described the serodiagnostic potential of I-PCR assays for an early diagnosis of TB based on the detection of potential mycobacterial antigens and circulating antibodies in body fluids of TB patients., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background and Objectives: Recently, third‐generation long‐read sequencing technology has been increasingly applied to the detection of various blood group systems. Because of its long read length and use of single‐molecule sequencing, it is capable of obtaining the sequences of blood group genes in their entirety as well as of distinguishing haplotypes. Therefore, here, we collected ABO blood group samples that were difficult to classify serologically and analysed the sequences of the coding regions of the ABO genes as well as the sequences upstream and downstream of the coding regions. Materials and Methods: Samples with ABO antigen typing and reverse serum typing discrepancies were screened in a total of 21 patients. All samples were subjected to serological testing and preliminary ABO genotyping (polymerase chain reaction with sequence‐specific primers [PCR‐SSP]), followed by single‐molecule real‐time (SMRT) sequencing to obtain complete ABO gene sequences. PCR sequence‐based typing (PCR‐SBT) was performed to validate the results. Results: Of the 21 samples, 15 had common ABO types, and 6 had rare ABO subtypes. One new allele, ABO*B.NEW (c.861C>T), and one allelic base recombination event was identified. Forty‐two haplotype sequences were obtained via SMRT sequencing with intronic single‐nucleotide variants (SNVs) specific to the ABO allele, and all of the exon region sequences were consistent with the PCR‐SBT results. Conclusion: SMRT sequencing is capable of accurately obtaining complete ABO gene sequences, distinguishing haplotypes and identifying allelic recombination. [ABSTRACT FROM AUTHOR]

Background: The Co-FriSero study describes a COVID-19 outbreak at the Friedrichroda hospital in Thuringia, Germany, with 185 beds and 404 employees, at the onset of the pandemic between March 30th, 2020, and April 13th, 2020. This study aimed to analyze potential sources of SARS-CoV-2 transmission amongst hospital employees. Methods: After the outbreak, a comprehensive follow-up was conducted through a questionnaire and a seroprevalence study using two different immunoassays for IgG detection and a third for discordant results. Results: PCR screenings confirmed SARS-CoV-2 infection in 25 of 229 employees, with an additional 7 detected through serology. Statistical analysis indicated that direct patient contact, exposure to high flow ventilation in non-isolated rooms, direct contact with colleagues, shared use of recreational rooms, and carpooling were associated with an increased infection risk. Conversely, contact with family and friends, public transportation, public events, and use of locker rooms were not associated with infection. Male gender showed a lower infection likelihood, independent of age and other risk factors. Conclusion: This study highlights the role of direct patient care and internal staff interactions in the spread of SARS-CoV-2 in the hospital setting. It suggests that non-traditional transmission routes like carpooling require consideration in pandemic preparedness. [ABSTRACT FROM AUTHOR]

Periodontitis is a chronic inflammatory destructive disease occurring in periodontal supporting tissues. Atherosclerosis(AS) is one of the most common cardiovascular diseases. Periodontitis can promote the development and progression of AS. Macrophage polarization is closely related to the development and progression of the above two diseases, respectively. The purpose of this animal study was to evaluate the effect of periodontitis on aortic lesions in atherosclerotic mice and the role of macrophage polarization in this process. 45 ApoE-/-male mice were randomly divided into three groups: control (NC), atherosclerosis (AS), and atherosclerosis with periodontitis (AS + PD). Micro CT, serological testing and pathological testing(hematoxylin–eosin staining, oil red O staining and Masson staining) were used for Evaluate the modeling situation. Immunohistochemistry(IHC) and immunofluorescence(IF) were performed to evaluate macrophage content and macrophage polarization in plaques. Cytokines associated with macrophage polarization were analyzed using quantitative real-time polymerase chain reaction(qRT-PCR) and enzyme-linked immunosorbent assay(Elisa). The expression of macrophages in plaques was sequentially elevated in the NC, AS, and AS + PD groups(P < 0.001). The expression of M1 and M1-related cytokines showed the same trend(P < 0.05). The expression of M2 and M2-related cytokines showed the opposite trend(P < 0.05). The rate of M1/M2 showed that AS + PD > AS > NC. Our preliminary data support that experimental periodontitis can increase the content of macrophage in aortic plaques to exacerbate AS. Meanwhile, experimental periodontitis can increase M1 macrophages, and decrease M2 macrophages, increasing M1/M2 in the plaque. [ABSTRACT FROM AUTHOR]

Objective: Brucellosis is an important zoonotic infection that causes multi-organ involvement, and the effectiveness of culture and serological tests used in the diagnosis of the disease is limited. In this study, it was aimed to investigate the effectiveness of Polymerase Chain Reaction (PCR) in the diagnosis of brucellosis. Material and Method: Sixty patients with positive Rose-Bengal Agglutination test and clinically suspected brucellosis and 60 control samples without these features were included in the study. The DNA of Brucella spp. was investigated in the samples by real-time PCR method. In the positive samples, the species identification was made, and virulance genes including omp19, wbkA, manA, mviN, urea and perA were studied with PCR. Results: Brucella spp. DNA was found to be positive in 19 of 32 patients in the patient group who were diagnosed with acute disease or relapse by clinical evaluation and serological tests, and no positive result was obtained in any of the control samples. Accordingly, the sensitivity and specificity of PCR were calculated as 59.3% and 100%, respectively. It was determined that Brucella melitensis was the infecting agent in all positive patients and carried all the virulence genes studied. Conclusion: Developing molecular methods and taking samples from the patient during the period of bacteremia may increase the effectiveness of PCR in the diagnosis of brucellosis. The identification of the pathogen as B. melitensis in all positive patients indicated that the transmission brucellosis to humans in our region was mainly from sheeps and goats, and appropriate control measures should be taken accordingly. [ABSTRACT FROM AUTHOR]

Introduction: Toxoplasmosis is a parasitism transmitted by Toxoplasma gondii, part of the TORCH complex, the most prevalent parasitism worldwide. It is asymptomatic in immunocompetent individuals but causes severe infections and developmental abnormalities in pregnant women, mainly affecting the central nervous system and the gastrointestinal system. Methods: In our prospective study, we analyzed cases of recent maternal Toxoplasma infections confirmed by serological testing between 1996 and 2020 at the Department of Obstetrics and Gynecology, Semmelweis University. Amniocentesis, followed by PCR, was performed in cases of recent infection confirmed by serological testing during pregnancy. After birth, a neonatological, microbiological, pediatric neurological and ophthalmological examination and a follow-up was carried out. Results: During the study period, a total of 238 cases of amniotic fluid Toxoplasma PCR testing due to Toxoplasma recent infection were performed. In terms of pregnancies, there were 219 deliveries and seven abortions. Twelve cases had no data available on the outcome of the pregnancy. In total, 133 cases of ultrasound abnormalities were detected during pregnancy, while in 105 cases, no abnormalities were detected on ultrasound examination. During amniocentesis, eight cases of Toxoplasma infection were revealed in amniotic fluid samples by PCR, and in 230 cases, the result was negative. Neonatal follow-up was performed in 139 cases, with no abnormalities during follow-up in 117 cases, and in 22 cases, there was a detectable complication that was likely to be related to Toxoplasma infection. In all 22 cases, amniotic fluid PCR Toxoplasma testing was negative. Conclusions: The most common ultrasound abnormalities involve the nervous system and the gastrointestinal system. In cases of suspicion, it is recommended to perform amniocentesis Toxoplasma PCR testing besides the indirect methods to help the pregnant woman decide whether to carry the pregnancy to term. During follow-up, a multidisciplinary team experienced in pregnancies complicated by toxoplasmosis must carry out the follow-up, care and subsequent development. [ABSTRACT FROM AUTHOR]

Background: The objective of this study is to analyze the transplacental transmission of SARS-CoV-2 antibodies, their persistence in newborns, the factors that may influence this transmission, and the protection these antibodies confer over time. Methods: This prospective cohort was conducted in a tertiary pediatric hospital in the Barcelona Metropolitan Region, Spain. It included neonates born to mothers who had SARS-CoV-2 infection during pregnancy or delivery between August 2020 and January 2022. We followed the recruited children for at least six months, and blood tests were performed to determine the presence of SARS-CoV-2 antibodies. Results: A total of 101 children were recruited. Among the serologies performed on children under three months of age, 44/82 were positive (53.7%). Newborns whose mothers presented more severe disease exhibited higher seropositivity odds (coefficient 9.747; p = 0.002). There were increased preterm deliveries when maternal infection occurred closer to the time of delivery. No severe SARS-CoV-2 infections were detected in children during the follow-up. Conclusions: Slightly more than half of the SARS-CoV-2 serologies performed in the first three months were positive. This appears to confer protection during early childhood. The severity of maternal infection is the most significant factor influencing the transmission of antibodies in children born to unvaccinated mothers. [ABSTRACT FROM AUTHOR]

Objectives In this feasibility study, we explored the combined use of circulating tumor human papillomavirus (HPV) DNA (ctHPVDNA) and HPV serology as diagnostic tests for HPV-associated oropharyngeal squamous cell carcinoma (OPSCC). Methods Among patients with research-banked serum or plasma at diagnosis, IgG antibodies to oncoproteins from HPV types 16, 18, 31, 33, 35, 45, 52, and 58 were detected with multiplex serology. Positivity for HPV 16 was defined based on detection of combinations of anti-E6, E1, E2, and E7 and for other high-risk types on detection of anti-E6 and anti-E7. Circulating tumor HPV DNA was detected by custom digital droplet polymerase chain reaction (ddPCR) assays for HPV types 16, 18, 33, 35, and 45. p16 immunohistochemistry and high-risk HPV RNA in situ hybridization (ISH) using a cocktail of 18 high-risk HPV types were performed on tissue. Results Of 75 patients, 67 (89.3%) were HPV-associated (p16 and HPV RNA ISH positive) and 8 (10.7%) were HPV-independent. All 8 HPV-independent patients were seronegative and negative for ctHPVDNA (100% specificity). Serology was positive in 53 (79.1%) of 67 HPV-associated patients, while ddPCR was positive for ctHPVDNA in 59 (88.6%) of 67 HPV-associated patients. Requiring both tests to be positive resulted in a sensitivity of 50 (74.6%) of 67 while combining assays (either positive) improved sensitivity to 62 (92.6%) of 67. Conclusions Compared to HPV RNA ISH, HPV serology and ctHPVDNA are sensitive and highly specific biomarkers for HPV-associated OPSCC at the time of presentation. [ABSTRACT FROM AUTHOR]

Background: On a global scale, approximately 350 million are affected by hepatitis B, and 71 million by hepatitis C. People in custody face elevated risks for these infections. The prevalence and risk factors in Iranian prisons are insufficiently documented. The principal objective of this study was to ascertain the prevalence of hepatitis B and C, coupled with the identification of pertinent influencing factors, within the confines of Zahedan central prison, situated in the southeastern region of Iran. Methods: In 2019, we conducted an analytical cross-sectional study involving 407 people in custody, using stratified random sampling. To definitively diagnose hepatitis C virus (HCV) infection (P < 0.05), a checklist developed by the researchers, along with enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) techniques, were employed. Results: This study comprised 406 participants (96.3% male) with a median age of 32 years (27-38). Approximately 62% were married, and a substantial proportion of the participants had low education levels (47%), unemployment (64%), and belonged to the Baloch ethnicity (64%). The overall prevalence of hepatitis C and B infections was 2.7% and 10.6%, respectively. Tattooing (adjusted odds ratio [AOR]: 2.07, 95% CI: 1.9-4.5) and marriage (AOR: 1.78, 95% CI: 1.05-3.04) were identified as risk factors for hepatitis B. Moreover, hepatitis C showed a statistically significant association with a family history of hepatitis B and C (AOR: 3.31, 95% CI: 3.93-24.64) and intravenous (IV) drug use (AOR: 7.01, 95% CI: 1.52-32.78) according to the multivariable logistic regression analysis. Conclusion: The prevalence of hepatitis B and C was higher among people in custody in Zahedan central prison. Consequently, targeted interventions are vital to address and reduce viral hepatitis burden in custodial settings. [ABSTRACT FROM AUTHOR]

Background: an accurate assessment of the immunity against SARS-CoV-2 can facilitate a better understanding and management of not only the recent coronavirus but similar pathogens as well. Objective: the aim of this study was to evaluate T-cell immunity with reference to antibody titers in a group of pediatric patients with autoimmune arthritides utilizing the widely known Interferon-γ Release Assay (IGRA). Materials and Methods: This study was conducted in the cohort of 55 children suffering from Juvenile Idiopathic Arthritis (JIA). This research analyzed the SARS-CoV-2 T-cell response measured by a specific quantitative IGRA, followed by a serological ELISA test measuring the presence and quantity of IgG, IgM, and IgA antibodies in serum. Results: The cellular response to SARS-CoV-2 measured by the IGRA test significantly correlated with the antibody titers, IgA (p < 0.00003, R = 0.537), IgG (p < 0.0001, R = 0.668), and IgG nucleocapsid protein (NCP) (p < 0.003, R = 0.0399), with no correlation with IgM levels. The antibody levels in patients receiving biological agents were significantly lower compared to the rest of the cohort (p = 0.0369), while traditional disease-modifying antirheumatic drugs had no such effect. Limitations: the main limitation of the research is the small sample size, mostly due to the specific cohort of patients and the lack of a healthy control. Conclusions: IGRA appears to be a viable tool in the accurate evaluation of T-cell responses to SARS-CoV-2, and serodiagnostics alone is not always sufficient in the assessment of immune responses. [ABSTRACT FROM AUTHOR]

Treatment of tularemia during pregnancy is challenging due to toxicity of standard treatment regimens. Here, we report a 31-year-old woman with glandular tularemia who was successfully treated with intravenous azithromycin. Follow-up examinations over a 6-month period showed a sustained response to treatment. She later gave birth to a healthy child. [ABSTRACT FROM AUTHOR]

Copyright of Gaceta Médica de Caracas is the property of Academia Nacional de Medicina and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objective The aim of this study was the development of an accurate and quantitative pyrosequence (PSQ) method for paternal RHD zygosity detection to help risk management of hemolytic disease of the fetus and newborn (HDFN). Methods Blood samples from 96 individuals were genotyped for RHD zygosity using pyrosequencing assay. To validate the accuracy of pyrosequencing results, all the samples were then detected by the mismatch polymerase chain reaction with sequence-specific primers (PCR-SSP) method and Sanger DNA sequencing. Serological tests were performed to assess RhD phenotypes. Results Serological results revealed that 36 cases were RhD-positive and 60 cases were RhD-negative. The concordance rate between pyrosequencing assay and mismatch PCR-SSP assay was 94.8% (91/96). There were 5 discordant results between pyrosequencing and the mismatch PCR-SSP assay. Sanger sequencing confirmed that the pyrosequencing assay correctly assigned zygosity for the 5 samples. Conclusion This DNA pyrosequencing method accurately detect RHD zygosity and will help risk management of pregnancies that are at risk of HDFN. [ABSTRACT FROM AUTHOR]

Objectives: This study aimed to assess the prevalence of blood‐borne infectious agents in healthy, client‐owned dogs from a blood donor population in Portugal and Spain, and to address the importance of a screening protocol on every donation. Materials and Methods: Client‐owned healthy dogs were tested before each donation on a veterinary blood bank. Blood samples from new potential donors, and from regular donors, were tested by real‐time polymerase chain reaction for Leishmania species, Ehrlichia species, Brucella species, Babesia species and Anaplasma species Serological tests were also performed for Leishmania species, Ehrlichia species and Dirofilaria immitis. All donors were tested for every infectious agent in each donation. Results: The study found that out of a total of 8036 donors and 35,120 samples tested, 3.9% of blood donors tested positive for at least one of the agents, with the most prevalent being Anaplasma species (2.1%). Serological tests also revealed positive results in 14.0% of donors, with the highest percentage for Leishmania species (7.7%). Moreover, the study found that 28.2% of positive results were from dogs with negative results in donations performed 3 to 12 months before, and 18.0% of positive results were recent infections. Clinical Significance: These findings indicate a high prevalence of infectious agents in seemingly healthy, selected dogs eligible to become blood donors in the Iberian Peninsula, highlighting the importance of regular testing on every donation. This study emphasises the importance of a regular screening protocol for every donation instead of annual testing, as is commonly performed in veterinary medicine. [ABSTRACT FROM AUTHOR]

Background Waning immunity after the coronavirus disease 2019 (COVID-19) vaccinations creates the constant need of boosters. Predicting individual responses to booster vaccines can help in its timely administration. We hypothesized that the humoral response to the first two doses of the BNT162b2 vaccine can predict the response to the booster vaccine. Methods A prospective cohort of hospital health care workers (HCW) that received three doses of the BNT162b2 vaccine. Participants completed serological tests at 1 and 6 months after the second vaccine dose and 1 month after the third. We analyzed predictive factors of antibody levels after the booster using multivariate regression analyses. Results From 289 eligible HCW, 89 (31%) completed the follow-up. Mean age was 48 (±10) and 46 (52%) had daily interaction with patients. The mean (±standard deviation) antibody level 1 month after the second vaccine was 223 (±59) AU/ml, and 31 (35%) had a rapid antibody decline (>50%) in 6 months. Low antibody levels 1 month after the second vaccine and a rapid antibody decline were independent predictors of low antibody levels after the booster vaccine. Conclusions The characteristics of the humoral response to COVID-19 vaccinations show promise in predicting the humoral response to the booster vaccines. [ABSTRACT FROM AUTHOR]

Chagas disease (CD), caused by Trypanosoma cruzi , is underdiagnosed in the United States. Improved screening strategies are needed, particularly for people at risk for life-threatening sequelae of CD, including people with human immunodeficiency virus (HIV, PWH). Here we report results of a CD screening strategy applied at a large HIV clinic serving an at-risk population. [ABSTRACT FROM AUTHOR]

Background and Aim: Cytomegalovirus (CMV) colitis is associated with negative outcomes in inflammatory bowel disease (IBD) and immunosuppressed cohorts and therefore requires timely recognition for appropriate management. We aimed to evaluate the diagnostic tools for CMV colitis and their associations with clinical outcomes. Methods: A retrospective cohort study of patients in a metropolitan health service with colonic samples analysed for CMV between 2012 and 2022, stratified into IBD and non‐IBD groups, was performed. The main outcome measures were the prevalence of positive and negative results for each CMV test, as well as need for colectomy, use of antiviral and hospital length of stay. Results: Five hundred eighty‐two biopsies from 418 patients were included; the median age was 36 years (interquartile range, 24–52 years) and 223 (53.3%) were men. Four hundred sixty‐one (79.2%) biopsies were from patients with IBD and 121 (20.8%) were from those without IBD. There were similar proportions of positive CMV histology (IBD 5.9% and non‐IBD 7.4%) and tissue CMV polymerase chain reaction (PCR) in the two groups (IBD 5.6% and non‐IBD 5.0%), but within each group, results were discordant. Positive CMV histology was significantly associated with need for colectomy in the IBD group, while positive tissue CMV PCR was not. Positive CMV histology, and tissue and serum CMV PCR were all significantly associated with antiviral use. Positive serum CMV PCR was significantly associated with colectomy. Conclusions: Histopathology remains the most predictive tool in assessing CMV colitis, while qualitative tissue CMV PCR was found to have limited utility. Quantitative serum CMV PCR may be useful but requires further evaluation. [ABSTRACT FROM AUTHOR]

Objectives: This study aimed to investigate the molecular mechanism of the Jk(a‐b‐) phenotype in a Chinese transfusion patient. Background: Many different mutation types relating to Jk(a‐b‐) phenotype have been reported. However, the splice‐site mutation is relatively rare and the related functional verification is lacking. Materials and Methods: In this study, the blood sample was collected from a transfusion patient with the Jk(a‐b‐) phenotype. Serotyping was performed using routine serological methods. The exons sequences and coding regions of the JK gene were amplified using polymerase chain reaction and directly sequenced. To perform a minigene splicing assay, the intronic mutation sequences were cloned into a pSPL3 splice reporting vector. The splicing reporter minigene assay was performed in HEK 293T cells. Results: The Jk(a‐b‐) phenotype of the blood sample was identified through serological testing. Sequencing results revealed that the sample had a novel homozygous splice‐site mutation JK*02N (NM_015865.7: c.663+3A>C). Further analysis, including cDNA sequencing and minigene splicing assay, confirmed that the novel splice‐site mutation resulted in exon skipping. Interestingly, different numbers of exons being skipped were obtained by the two methods. Conclusion: This study revealed a novel homozygous splicing‐site mutation associated with the Jk(a‐b‐) phenotype in Chinese population. Our results emphasise the importance of the in vitro functional method minigene splicing assay, while also acknowledging its potential limitations when compared to cDNA sequencing. [ABSTRACT FROM AUTHOR]

Introduction: This study standardized a semi-quantitative dot blotting assay (DB) and a quantitative real-time polymerase chain reaction (qPCR) to detect specific antibodies for Paracoccidioides brasiliensis and its DNA in PCM patients. Methodology: We evaluated 42 confirmed PCM patients upon admission using a serological double agar gel immunodiffusion test (DID), DB, and molecular tests (qPCR in total blood). The control groups included 42 healthy individuals and 37 patients with other infectious diseases. The serological progress during treatment was evaluated in eight patients, and there was a relapse diagnosis in ten patients using the Pb B.339 strain antigen. The cut-off points for the serological tests were determined by a receiver operator characteristic curve. Results: The DB and DID tests showed similar accuracy, but the DB identified lower antibody concentrations. Cross-reactions were absent in the DB assay. In the relapse diagnoses, DB exhibited much higher sensitivity (90%) than DID (30%). Conclusions: A DB assay is easier and faster than a DID test to be performed; DB and DID tests show the same accuracy, while blood qPCR is not recommended in the diagnosis at the time of admission; cross-reactions were not observed with other systemic diseases; DB and DID tests are useful for treatment monitoring PCM patients; and a DB assay is the choice for diagnosing relapse. These findings support the introduction of semi-quantitative DB assays in clinical laboratories. [ABSTRACT FROM AUTHOR]

Background: Healthcare workers (HCWs) faced substantial risk of infection during the COVID-19 outbreak. This study aims to determine the prevalence of anti-SARS-CoV-2 antibodies in a cross-sectional sample of HCWs as well as risk factors associated with exposure to SARS-CoV-2. Methods: The study was conducted between March and May 2021 at the American University of Beirut Medical Center (AUBMC), a tertiary hospital located in Lebanon. Socio-demographic and clinical data, as well as data on exposure, PCR results, PPE adherence, and vaccination status, were collected using an online questionnaire. Sera were also collected to determine seropositivity using commercially available enzyme-linked immunoassay (ELISA) targeting the spike (S) and the nucleocapsid proteins (NCP) of SARS-CoV-2. Findings: Among 92 recruited HCWs, 72.3% received PPE training, more than 70% were adherent to using appropriate PPEs, and around 80% were vaccinated. Nurses in this study population were at higher risk of exposure compared to medical doctors, technicians, and other HCWs. Among the HCWs who performed a PCR test, 28.6% were infected with SARs-CoV-2 with workplace exposure not associated with COVID-19 infection. All vaccinated HCWs were seropositive for anti-S IgG with high titer (≥384 BAU/mL), with a significantly higher median anti-S IgG titer compared to unvaccinated HCWs with previous infection (384 vs. 140.1 BAU/mL; p =.0043). Conclusions: Our study highlights the importance of implementing strict infection control policies among HCWs and deploying an effective COVID-19 vaccination strategy. More studies are needed in Lebanon to assess risk factors of SARS-CoV-2 exposure in the workplace. [ABSTRACT FROM AUTHOR]

Background: Malakoplakia is a rare chronic granulomatous disease characterized by the presence of Michaelis-Gutmann bodies (MGBs) within histiocytic aggregates. It predominantly affects immunocompromised individuals, including those living with Human Immunodeficiency Virus (HIV). Case Presentation: We present a unique case of bifocal malakoplakia in a 49-year-old man, previously with Coronavirus disease 2019 (COVID-19) and HIV positive, presented with respiratory symptoms, weight loss, and lymphadenopathy. He had various infections including Non-Tuberculous Mycobacteria (NTM), Cytomegalovirus (CMV), and Candida, with evolving lung and gastrointestinal issues. Despite treatment attempts, he deteriorated due to respiratory distress, multi-organ failure, and coagulopathy, leading to his unfortunate demise. Conclusion: This report presents a distinctive and complex case of malakoplakia in an HIV-positive patient, a rare inflammatory disorder originally described by Michaelis and Gutmann in 1902. The hallmark Michaelis-Gutmann organisms were observed, confirming the diagnosis. While typically affecting the urinary tract, this case demonstrates the exceptional ability of malakoplakia to manifest in various organ systems, including pulmonary, gastrointestinal, and more. Although Escherichia coli is a prevalent associated pathogen, the exact cause remains elusive. Treatment, often involving surgical excision and antibiotic therapy, underscores the challenging nature of managing this condition in immunocompromised individuals. [ABSTRACT FROM AUTHOR]

The article discusses research which examined the use of SARS-CoV-2 serology to assess risk of SARS-CoV-2 breakthrough infection (BTI) after COVID-19 vaccination during the omicron surge. The study measured serology and identified BTI from weekly saliva self-collection. It describes the demographics and clinical characteristics prior to index date, COVID-19 vaccination, and SARS-CoV-2 BTI by anti-nucleocapsid and anti-spike protein receptor-binding domain serology results.

Background: Shigella spp., which are facultative anaerobic bacilli within the Enterobacteriaceae family, present a significant public health burden due to their role as prominent contributors to diarrheal diseases worldwide. A molecular analysis can facilitate the identification and assessment of outbreaks involving this bacterium. So, we aimed to investigate the antibiotic susceptibility pattern and clonal relatedness of clinical Shigella spp. isolates obtained from patients with diarrhea in Hormozgan province, South of Iran. Methods: From 2019 to 2021, a cross-sectional investigation was conducted on 448 stool samples obtained from patients who were experiencing diarrhea, in the southern region of Iran. Shigella spp. isolates were identified based on biochemical and serological tests. All Shigella species were verified using species-specific polymerase chain reaction (PCR), followed by susceptibility testing to antimicrobial agents. Subsequently, genotyping of all Shigella species was conducted using ERIC-PCR. Results: Out of a total of 448 stool samples, the presence of Shigella was detected in 62 cases, accounting for a prevalence rate of 13.84%. Among the identified isolates, the majority were attributed to S. flexneri, representing 53.23% of the cases. This was followed by S. sonnei at 24.19% and S. boydii at 22.58%. Notably, no instances of S. dysenteriae were found. The highest prevalence of Shigella isolates was observed in infants and children under the age of five. A significant proportion of the identified isolates demonstrated resistance to various antibiotics. Specifically, high resistance rates were noted for ampicillin (90.78%), piperacillin–tazobactam (87.1%), cefixime (83.87%), trimethoprim–sulfamethoxazole (83.87%), cefotaxime (82.26%), and ceftriaxone (80.65%). In addition, a substantial number (87.1%) of the isolates exhibited a multidrug-resistant (MDR) phenotype. Using the ERIC-PCR method, a total of 11 clusters and 6 distinct single types were identified among all the Shigella isolates. Conclusion: A notable occurrence of antibiotic-resistant Shigella species has been noted, with multi-drug resistant (MDR) strains presenting an increasing challenge for treating shigellosis worldwide, and this includes Iran. Techniques such as ERIC-PCR are useful for assessing the genetic variation and connections between Shigella strains, which indirectly contributes to understanding antimicrobial resistance patterns. Further research is needed to explore the specific correlation between resistance genes and ERIC genotyping patterns in Shigella strains. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is considered as one of the leading causes of food-intoxication and on the other hand E. coli and particularly, Shiga toxin (ST) producing Escherichia coli (STEC) have emerged as important food-borne enteropathogens frequently associating serious to fatal disorders in humans and both species have shown to be resistant to a wide spectrum of antibiotics. The current study was performed to determine the prevalence of two common food-borne pathogens: Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli ) in white soft cheese being one of the most popular dairy products in Egypt. A total 150 samples of soft cheese were purchased from the market and were microbiologically tested. The S. aureus isolates were identified according to recommended biochemical tests, and polymerase chain reaction (PCR) targeting the nuc gene, as a species-specific marker gene for S. aureus. Meanwhile, E. coli isolates were identified based on biochemical tests, serological tests, haemolytic activity, and multiplex PCR. This has been done to assess capability of producing STs based on targeting stx1 and stx2 genes. Result revealed that S. aureus and E. coli were observed in 66.66% and 36% of samples, respectively. The antimicrobial resistance of isolates was phenotypically investigated against eleven different antibiotics and results showed the presence of a variable multidrug-resistance among isolates against selected antibiotics. These findings highlighted the importance of periodical screening of food-borne pathogens in artisanal products due to lack of strict hygienic measures in markets; the reason that facilitate the contamination of such artisanal products by several food-borne pathogens. [ABSTRACT FROM AUTHOR]

Laboratory investigations can play a significant role in the diagnosis and decision-making of infectious uveitis. Though direct demonstration of the infective organism remains the gold standard of diagnosis, it is not always possible with ocular tissues. Recent advancements in molecular techniques have made it possible to overcome these limitations and to identify the genomic DNA of pathogens associated with infectious uveitis. Techniques such as next-generation sequencing can analyze all DNA-based lifeforms, regardless of whether they are bacteria, fungi, viruses, or parasites and have been used in the laboratory diagnosis of intraocular inflammation. On the other hand, serological tests, though they dominate the diagnostic landscape of various infectious etiologies in uveitis in routine clinical practice, have varied specificities and sensitivities in different infectious uveitis. In this review, we focus on various methods of laboratory diagnosis of infectious uveitis and discuss the recent advances in molecular diagnosis and their role in various infectious clinical entities. [ABSTRACT FROM AUTHOR]

Background Vaccination may control the coronavirus disease 2019 (COVID-19) pandemic, including in nursing homes where many high-risk people live. We conducted extensive outbreak investigations. Methods We studied an outbreak at a nursing home in Switzerland, where the uptake of messenger RNA vaccines against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was 82% among residents as of 21 January 2021. After diagnosis of COVID-19 in a vaccinated symptomatic healthcare worker (HCW) on 22 February, we performed outbreak investigations in house A (47 residents; 37 HCWs), using SARS-CoV-2–specific polymerase chain reaction testing of nasopharyngeal swab samples. We performed whole-genome sequencing of SARS-CoV-2 and serological analyses. Results We identified 17 individuals with positive polymerase chain reaction results, 10 residents (5 vaccinated) and 7 HCWs (3 vaccinated). The median age (interquartile range) was 86 (70–90) years among residents and 49 (29–59) years among HCWs. Of the 5 vaccinated residents, 3 had mild disease and 2 had no symptoms, whereas all 5 unvaccinated residents had mild to severe disease, and 2 died. Vaccine effectiveness for the prevention of infection among residents was 73.0% (95% confidence interval, 24.7%–90.1%). The 12 available genomes were all alpha variants. Neutralizing titers were significantly higher in vaccinated individuals on reexposure (>1 week after diagnosis) than in vaccinated, unexposed HCWs (P  = .01). Transmission networks indicated 4 likely or possible transmissions from vaccinated to other individuals and 12 transmission events from unvaccinated individuals. Conclusions COVID-19 outbreaks can occur in nursing homes, including transmission from vaccinated persons to others. Outbreaks might occur silently, underlining the need for continued testing and basic infection control measures in these high-risk settings. [ABSTRACT FROM AUTHOR]

Background and Objectives: The extremely rare Rhnull phenotype is characterized by the absence of all Rh antigens on erythrocytes. It is divided into the regulator and amorph types based on the underlying genetic background. The more common regulator type depends on critical variants silencing RHAG, which encodes RhAG glycoprotein, necessary for RhD/RhCE expression. Rhnull cells have altered expression of glycophorin B and LW glycoprotein. Materials and Methods: Four unrelated Rhnull individuals were investigated. Serological testing was performed according to standard blood bank practice. RHD/RHCE and S/s allele‐specific Polymerase chain reaction (PCR) genotyping was done on genomic DNA using in‐house PCR assays. RHAG, and in some cases also RHD/RHCE, were sequenced. Initial s phenotyping results triggered additional serological investigation. Results: Anti‐Rh29 was identified in all four individuals. Extended typing with anti‐S and anti‐s showed that the three samples predicted to type as s+ failed to react with 2 of 5 anti‐s. Sequence analysis of all 10 RHAG exons and the immediate intron/exon boundaries revealed a single nucleotide variant in the 3′‐end of intron 6, c.946 −2a>g in all samples. RHD/RHCE showed no alterations. Conclusion: A novel Nordic Rhnull allele was identified. In addition, it was shown that s+ Rhnull red blood cells are not only U− but also have qualitative changes in their s antigen expression. [ABSTRACT FROM AUTHOR]

Background Tick-borne diseases have grown in incidence over recent decades. As a result, diagnostic testing has become more common, often performed as broad antibody-based panels for multiple tick-transmitted pathogens. Rocky Mountain spotted fever (RMSF) is rare in our region yet may cause severe morbidity, leading to diagnostic screening in low-risk patients. We sought to describe trends in RMSF diagnostic testing, rate of IgG seropositivity, and clinical features of those tested. Methods We performed a retrospective chart review of patients ≤21 years old undergoing testing for RMSF and/or with an ICD-9/10 code for RMSF. Patients were categorized by infection likelihood based on clinical and laboratory criteria adapted from Centers for Disease Control and Prevention's (CDC) case definition of spotted fever rickettsioses. Clinical data were collected and analyzed with descriptive statistics. Results One hundred and seventy patients were included. 5.8% met CDC criteria for rickettsial infection, 6.5% had an elevated IgG titer but lacked suggestive symptoms, and 87.6% had a negative IgG titer. Many patients tested were unlikely to have RMSF, including 50% lacking fever, 20% lacking any RMSF "classic triad" symptoms, 13% without acute illness, and 22% tested during months with low tick activity. Convalescent serology was performed in 7.6% of patients and none underwent Rickettsia rickettsii polymerase chain reaction (PCR) testing. Conclusions Diagnostic testing was frequently performed in patients unlikely to have RMSF. We identified many opportunities for improving test utilization. Reserving testing for those with higher pretest probability, performing convalescent serology, and utilizing PCR may improve the accuracy of RMSF diagnosis and reduce clinical challenges stemming from inappropriate testing. [ABSTRACT FROM AUTHOR]

BACKGROUND AND OBJECTIVE: A number of glycophorin variant phenotypes or hybrid glycophorin variants of the MNS blood group system bear multiple immunogenic antigens such as Mia, Mur, and MUT. In the East and Southeast Asian populations, glycoprotein (GP.) Mur is the most common glycophorin variant phenotype expressing those three immunogens. The aim of this study was to detect MNS system glycophorin variant phenotypes (GP. Mur, GP. Hop, GP. Bun, GP. HF, and GP. Hut) among Malaysian blood donors. MATERIALS AND METHODS: In this cross-sectional study, 144 blood donors were selected under stratified random sampling. The deoxyribonucleic acid was extracted from whole blood samples, followed by a polymerase chain reaction assay. Sanger sequencing was used to identify the specific MNS variants and then validated by a serological crossmatch with known anti-Mur and anti-MUT. RESULTS: GP. Mur was identified among Malaysian blood donors with a prevalence of 6.94%, and no other variants of the MNS system were found. CONCLUSION: The present study substantiates that GP. Mur is the main variant of the MNS system glycophorin (B-A-B) hybrid in Malaysian blood donors. GP. Mur-negative red blood cells must therefore be considered in the current transfusion policy in order to prevent alloimmunization and immune-mediated transfusion reactions, particularly in transfusion-dependent patients. [ABSTRACT FROM AUTHOR]

Introduction:Babesia is a tick-borne intraerythrocytic parasite that is globally ubiquitous, yet understudied. Several species of Babesia have been shown to be transfusion-transmissible. Babesia has been reported in blood donors, animals, and ticks in the Tyrol (Western Austria), and regional cases of human babesiosis have been described. We sought to characterize the risk of Babesia to the local blood supply. Methods: Prospective molecular testing was performed on blood donors who presented to regional, mobile blood collection drives in the Tyrol, Austria (27 May to October 4, 2021). Testing was conducted using the cobas® Babesia assay (Roche Molecular Systems, Inc.), a commercial PCR assay approved for blood donor screening that is capable of detecting the 4 primary species causing human babesiosis (i.e., B. microti, B. divergens, B. duncani, and B. venatorum). A confirmatory algorithm to manage initial PCR-reactive samples was developed, as were procedures for donor and product management. Results: A total of 7,972 donors were enrolled and screened; 4,311 (54.1%) were male, with a median age of 47 years (IQR = 34–55). No positive cases of Babesia were detected, corresponding with an overall prevalence of 0.00% (95% CI: 0.00%, 0.05%). Discussion: The findings suggest that the prevalence of Babesia is low in Austrian blood donors residing in the Tyrol, even during months of peak tick exposure. Although one cannot conclude the absence of Babesia in this population given the limited sample size, the findings suggest that the regional risk of transfusion-transmitted babesiosis is low. [ABSTRACT FROM AUTHOR]

Background: Lua and Lub are inherited as codominant allelic characters resulting from a single nucleotide variant (SNV) of the basal cell adhesion molecule (BCAM) gene. Red cells of the dominantly inherited suppressor of the Lutheran antigens In(Lu) phenotypically appear as Lu(a−b–) by the haemagglutination test. In(Lu) resulted from heterozygosity for mutations within the erythroid-specific Krüppel-like factor 1 (KLF1) gene. This study aimed to determine the frequency of the Lu(a) and Lu(b) phenotypes and genotypes and genetic variants of the distinct In(Lu) among Thai blood donors. Material and Methods: Samples from 334 Thai donors were phenotyped with anti-Lua and anti-Lub. These DNA samples and an additional 1,370 donor DNA samples with unknown Lu(a)/Lu(b) phenotypes were genotyped using an in-house PCR-SSP. In the case of the three Lu(a–b–) donors, the BCAM and KLF1 genes were analysed by PCR and sequencing. Results: A total of 331 of the 334 donors were Lu(a–b+), while the other observed phenotype, appearing as Lu(a–b–), was found among three donors. Of those three Lu(a–b–) donors with the LU*02/02 genotype, we identified KLF1 variant alleles, consisting of two variants: c.[304T>C, 1001C>G] and c.[304T>C, 519_525dupCGGCGCC], leading to the In(Lu) phenotype, and one homozygous variant (c.304T>C) mutation. Also, only one Thai donor was genotyped as LU*01/02, confirmed by serology test and DNA sequencing. Conclusion: In this study, we identified KLF1 variants to be included in Lutheran typing analysis in Thai populations. Therefore, the application of genotyping and phenotyping methods has simultaneously been in use to screen and confirm the rare Lu(a+) and In(Lu) phenotypes. [ABSTRACT FROM AUTHOR]

Objective Estimating severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) seroprevalence is an important part of the public health approach to coronavirus disease 2019 (COVID-19) understanding and containment. This is particularly relevant to an obstetric population because of implications in the management of the pregnant host, care of the newborn, and disease progression within the community. Study Design A cross-sectional seroprevalence study was performed in four Department of Health Palm Beach County clinics from June 29, 2020, to August 5, 2020. Samples were collected from asymptomatic antepartum and postpartum participants. A web-based surveillance system was used to identify subsequent antibody or polymerase chain reaction (PCR) testing encounters. Results A total of 163 of 618 subjects were seropositive (26.4%). Racial makeup was white 2.5%, black 19.0%, and Hispanic 78.5%. Positive serology was seen in 16.0, 35.6, and 30.1% of first, second, and third trimesters, respectively; 18.4% were positive postpartum. Only four patients voluntarily reported PCR positivity prior to antibody testing. Six home zip codes accounted for the majority (68.1%) of positive results. Thirty-two patients had repeat serology (65.6% positive and 34.4% negative). Of the 163 subjects, 65 underwent later PCR testing with 92% negative for SAR-CoV-2. Conclusion Almost one in four subjects had serologic evidence of previous SARS-CoV-2 infection. These very high seroprevalence rates have not been previously reported and highlight the concern for health disparities in the United States. Most were asymptomatic and without a history for SARS-CoV-2 exposure. There was a loss of seropositivity in a significant number of subjects, raising concern for risk of reinfection, inadequate transplacental antibody transfer, and subsequent limited passive protection to the newborn. These seroprevalence data will also allow for better newborn follow-up of unanticipated consequences of COVID-19 infection in pregnancy. Key Points SAR-CoV-2 seroprevalence is disproportionately high in this obstetric population The majority of subjects who tested positive for SAR-CoV-2 were asymptomatic for COVID-19 infection Estimating seroprevalence is an important public health approach to disease surveillance and understanding [ABSTRACT FROM AUTHOR]

• A disease occurrence characterized by fever and rash was monitored in China. • The etiological and epidemiological investigation was initiated. • We proved that a novel species of genus Ehrlichia was its pathogen. • We named the new species as " Candidatus Ehrlichia erythraense". From March to June 2021, the reported number of clinically diagnosed endemic typhus in Anhui and Hubei provinces of China nearly increased four-fold compared with the monthly average numbers in last 5 years. An etiological and epidemiological investigation was initiated. The clinical specimens from the reported patients and the potential vector ticks were collected for molecular and serological detection, as well as cell culturing assay to identify the potential pathogen. Polymerase chain reaction and sequence analysis of rrs and groEL showed that the pathogen from these patients was Ehrlichia sp., isolated from Haemaphysalis longicornis attached to these patients. The phylogenetic analysis based on 39 Ehrlichia genomes suggested that it should be taxonomically classified as a novel species, tentatively named " Candidatus Ehrlichia erythraense". A total of 19 of 106 cases were confirmed as Candidatus Ehrlichia erythraense infections by polymerase chain reaction, sequencing, and/or serological tests. The most frequent symptoms were fever (100%), rashes (100%), asthenia (100%), anorexia (100%), and myalgia (79%). The occurrence of the disease presenting with fever and rashes in Anhui and Hubei provinces was caused by a novel species of the genus Ehrlichia ; physicians need to be aware of this newly-discovered pathogen to ensure appropriate testing, treatment, and regional surveillance. [ABSTRACT FROM AUTHOR]

Nucleic acid amplification tests (NAATs), including polymerase chain reaction (PCR) assays, are more sensitive for the detection of SARS-CoV-2 than rapid antigen tests (RATS), and are the gold standard for diagnosis of acute COVID-19. However NAATs can remain positive for weeks following infection due to low-level shedding of non-viable viral fragments. RATs (in particular self-testing) are the mainstay of COVID-19 diagnosis due to their convenience, speed and high specificity. The sensitivity of RATs is highest within seven days of symptom onset. A negative RAT result may warrant a NAAT or repeat RAT for confirmation. The presence of spike antibodies is consistent with either vaccination or infection. Nucleocapsid antibodies suggest a previous infection. Serological tests measuring neutralising antibodies that infer immunity are not readily available. [ABSTRACT FROM AUTHOR]

Aims: Bronchial asthma is a life-threatening disease with a multifactorial etiology. It was shown that immunological dysregulations play an essential role in the exacerbation of the patient's attacks. Tolllike receptors act as a bridge in the transmission of different signals associated with the regulation of immune responses. According to numerous research, TLR2 causes different outcomes in asthma. TLR2 and TLR4 overexpression seems to predispose to an increase in the frequency of dyspneic attacks. Therefore, this study aimed to learn more about the function of TLR2 and TLR4 and investigate their underlying association with allergic airway inflammation in asthmatic patients. Malerials & Methods: In this case-control study conducted at outpatient clinics in a Specialized Allergy Center in Basrah City, South of Iraq, 70 asthmatic patients and 70 healthy individuals were investigated. 4ml of venous blood was drawn and divided into two parts: 2ml was used for RNA extraction and qRT-PCR, and another 2ml was used for the serological tests. Data analysis was done by SPSS 26 software. Findings: The mean serum levels of TLR2 and TLR4 were significantly higher in the asthmatic group (28.33±13.00ng/ml and 46.30±20.32ng/ml, respectively) than in the control group (7.33±3.73ng/ml and 26.28±14.32ng/ml, respectively). Also, the expression level of TLR2 and TLR4 was significantly higher in the asthmatic group (p<0.05). Conclusion: TLR2 and TLR4 are increased in patients with bronchial asthma compared to healthy individuals. Elevated levels of both biomarkers may act as a trigger for increased secretion of other cytokines, leading to exacerbation of asthmatic signs and symptoms. [ABSTRACT FROM AUTHOR]

Background: Hemolytic uremic syndrome (HUS) is characterized by microangiopathic hemolysis, thrombocytopenia, and thrombus formation leading to tissue injury. HUS is classified according to its etiology as post-diarrheal or atypical HUS. Differential diagnosis of both entities continues to be a challenge for pediatric physicians. Methods: The aim was to improve the rapid etiological diagnosis of post-diarrheal HUS cases based on the detection of Shiga toxin (Stx)-producing Escherichia coli (STEC) infection by screening of stx1/stx2 and rfbO157 in cultured stools by multiplex PCR, and the additional detection of anti-lipopolysaccharide (anti-LPS) O157, O145, and O121 antibodies by Glyco-iELISA test. In addition, we studied patients' relatives to detect circulating pathogenic strains that could contribute to HUS diagnosis and/or lead to the implementation of measures to prevent dissemination of familial outbreaks. This study describes the diagnosis of 31 HUS patients admitted to Hospital Municipal de Niños Prof Dr Ramón Exeni during the 2017–2020 period. Results: Stool PCR confirmed the diagnosis of STEC associated with HUS in 38.7% of patients (12/31), while anti-LPS serology did in 88.9% (24/27). In those patients in which both methods were carried out (n = 27), a strong association between the results obtained was found. We found that 30.4% of HUS patients had at least one relative positive for STEC. Conclusions: We could identify 96.3% (26/27) of HUS cases as secondary to STEC infections when both methods (genotyping and serology) were used. The results demonstrated a high circulation of STEC in HUS families and the prevalence of the STEC O157 serotype (83%) in our pediatric cohort. A higher-resolution version of the Graphical abstract is available as Supplementary information. [ABSTRACT FROM AUTHOR]

Expanding a previous study of the immune response to SARS-CoV-2 in 10 New Jersey long-term care facilities (LTCFs) during the first wave of the pandemic, this study characterized the neutralizing antibody (NAb) response to infection and vaccination among residents and staff. Sera from the original study were tested using the semi-quantitative enzyme-linked immunosorbent cPass neutralization-antibody detection assay. Almost all residents (97.8%) and staff (98.1%) who were positive for IgG S antibody to the spike protein were positive for NAb. In non-vaccinated subjects with a history of infection (positive polymerase chain reaction (PCR) or antigen test), the distribution of mean intervals from infection to serology date was not significantly different for S antibody positives versus negatives. More than 80% of both were positive at 10 months. Similarly, the mean NAb titer for residents and staff was not associated with interval from PCR/antigen positive to serology date, F = 0.1.01, Pr > F = 0.4269 and F = 0.77, Pr > F = 0.6548 respectively. Titers remained high as the interval reached 10 months. In vaccinees who had no history of infection, the NAb titer was near the test maximum when the serum was drawn seven or more days after the second vaccine dose. In staff the mean NAb titer increased significantly as the vaccine number increased from one to two doses, F = 11.69, Pr > F < 0.0001. NAb titers to SARS-CoV-2 in residents and staff of LTCFs were consistently high 10 months after infection and after two doses of vaccine. Ongoing study is needed to determine whether this antibody provides protection as the virus continues to mutate. [ABSTRACT FROM AUTHOR]

Background: Syphilis is a sexually transmitted infection (STI) caused by the pathogen Treponema pallidum. Its incidence is increasing in our country, especially among men who have sex with men (MSM). Serological tests are still the most widely used technique for diagnosis. The need for an early diagnosis has prompted the introduction of fast techniques, such as Treponema pallidum detection by polymerase chain reaction (PCR) on mucocutaneous samples. The objective of this work is to analyse the sensitivity of this technique in a series of patients diagnosed with syphilis at our centre. Methods: Retrospective review of all cases diagnosed with syphilis at our centre between May 2017 and May 2021. Results: A total of 203 cases of syphilis were diagnosed with serologic tests: 33% were primary syphilis and 53.1% secondary syphilis. PCR for Treponema pallidum was performed in 117 (57,6%) cases. The sensitivity was highest (95,2%) when performed on samples from mucocutaneous ulcers in primary syphilis. This value decreased to 69,4% in secondary syphilis, although there were variations between the types of samples. Conclusions: The PCR test has a high diagnostic value when performed on ulcer exudates in patients with primary syphilis. Its most relevant advantages in clinical practice are the possibility of an early diagnosis before serological tests during the window period, the ability to confirm reinfections in patients with persistent positivity of reaginic antibodies and a history of treated syphilis. Nevertheless, given that a negative PCR test may not rule out infection by Treponema pallidum, serologic tests are still necessary for everyday practice. [ABSTRACT FROM AUTHOR]

BACKGROUND: In multi-transfused thalassemia patients, serological phenotyping fails to test patient's actual blood group antigen profile due to the presence of donor red blood cell (RBC) in the circulation. This limitation of serological tests can be overcome by genotype determination using the polymerase chain reaction (PCR)-based methods. The aim of this study is to compare the serological phenotyping of Kell, Kidd, and Duffy blood group systems with molecular genotyping in the normal blood donors and multi-transfused thalassaemia patients. MATERIALS AND METHODS: Blood samples from 100 normal blood donors and 50 thalassemia patients were tested using standard serological techniques and PCR-based methods for Kell (K/k), Kidd (Jka/Jkb), and Duffy (Fya/Fyb) blood group systems. The results were compared for concordance. RESULTS: Genotyping and phenotyping results were 100% concordant for normal blood donors whereas those for thalassemia patients showed 24% discordance. The frequency of alloimmunization in thalassemia patients was 8%. The results of genotyping were used to provide Kell, Kidd, and Duffy matched blood for transfusion therapy to thalassemia patients. CONCLUSION: The actual antigen profile in multitransfused thalassaemia patients can be reliably determined using genotyping. This would benefit in providing better antigen matched transfusion therapy to such patients hence reducing the rate of alloimmunization. [ABSTRACT FROM AUTHOR]

Background: As one of the most widespread sexually transmitted infections, Herpes Simplex Virus (HSVs) globally account for 60-95% of persistent infections in adults. This infection is prevalent in women of gestational age and is likely to be transmitted from the infected mother to her neonate. Additionally, it gives rise to devastating complications in neonates. This study was designed to estimate the molecular and serological prevalence of HSV-1 and 2 in pregnant women of Gorgan city, North East of Iran. Methods: Vaginal secretions and blood specimens of 315 pregnant women referred to an educational hospital in the North east of Iran were tested for HSV-1 and HSV-2 using multiplex PCR and ELISA assays. Chi-Square test was utilized to evaluate the association of qualitative variables and the level of significance was set at p≤0.05. Moreover, statistical analysis was performed using SPSS V.19.0. Results: HSV-1 and HSV-2 DNA was detected in 5.7% and 8.3% of participants, respectively. Given the serological analyses of total HSV-1 and HSV-2 antibodies, 92.7% (239/315) of patients were IgG positive and 5.4% (17/315) were IgM positive. Conclusion: The rate of HSV-1 and 2 in the present study was lower than that reported by World Health Organization (WHO). This study emphasizes the conduction of further investigations on HSVs since these viruses are probably playing significant role in sexually transmitted infections. [ABSTRACT FROM AUTHOR]

Background: Studies in different communities have shown significant differences in IgG antibody titers in the time period after the first and second doses of the vaccines. This study aimed to serologically evaluate the IgG anti-spike antibody titer five months after injection of the second COVID-19 vaccine in healthcare workers. Materials and method: This study was performed in healthcare personnel for whom five months had passed since their second anti-Covid-19 vaccination. The level of IgG antibody against SARS-CoV-2 spike protein was measured by ELISA. Healthcare workers in Mofid Children's hospital received three brands of vaccines: Sputnik V, Sinopharm, and AstraZeneca. Results: The mean titer of anti-spike IgG was 4.3±2.29 units. The percentage of positive cases of the antibody was estimated to be 96.4%. The titer of anti-spike IgG antibody was dependent on both the occupational area and a positive history of Covid-19 disease. Conclusion: About 96.4% of the staff vaccinated against Covid-19 had a high titer of anti-spike IgG antibody even five months after inoculation of the second dose. The field of occupational can affect the level of antibody present [ABSTRACT FROM AUTHOR]

Background and Objectives: Polar lipids and the 16S rRNA gene have a significant role in taxonomic characteristics. The study’s objective is to determine the potential relationship between simple migration distances (mm) of polar lipids and Salmonella’s 16S rRNA gene similarity. Materials and Methods: Based on 16S RNA sequences and simple thin‑layer chromatography migration distances (mm) of polar lipids, allowed to compare the various Salmonellae species, and apply it to examine the variability and estimate bacterial similarities. Results: The chromatography migration distance analysis revealed 3–4 spots of polar lipids. The polar lipids revealed two denser spots, the most abundant lipids, between 10 and 28 mm. The other two low‑density spots of migration distance ranged in size from 23 to 25 mm. Between 99.4% and 100% of the three Salmonella isolates and other Salmonella species exhibited 16S rRNA similarities. One strain had a similarity of 98.9%. These findings demonstrated the nearly identical 16S rRNA gene sequences and polar lipids profile of the isolates. Conclusions: This study has concluded that all Salmonella species share similarities in both the polar lipid profiles and the 16S rRNA genes. The study validates the utility of coupling of polar lipids and 16S rRNA gene sequencing as useful tools for taxonomic differentiation and epidemiological tracing of Salmonella. [ABSTRACT FROM AUTHOR]

Leptospirosis is a serious zoonotic infection and the most prevalence disease is in the tropical and subtropical region. The definitive diagnosis of Leptospirosis, caused by spirochetes of the genus Leptospira infection is already using culture methods, serological tests such as the microscopic agglutination test (MAT) and molecular detection methods (PCR) are possible. In this study, we used multiplex PCR method for detection of pathogenic and non - pathogenic Leptospira based on lipL32 and 16S rRNA genes. All serovars were obtained from the Leptospira Reference Laboratory of Microbiology Department, Razi Vaccine and Serum Research Institute, Karaj, Iran. The PCR product for the lipL32 and 16S rRNA genes was 272 bp and 240 bp respectively. The sensitivity amplification for the multiplex assay was 10-6 pg / µl for 16S rRNA gene and 10-4 pg / µl for lipL32 gene. The sensitivity for multiplex PCR was 10-3 pg / µl. The results supported the idea that multiplex PCR can be used to detect Leptospira samples. This method was also able to differentiate between saprophytic and pathogenic leptospires and was able to do so much easily than conventional methodologies. Due to the slow growth of Leptospira and the importance of time in diagnosis, molecular methods such as PCR are suggested. [ABSTRACT FROM AUTHOR]

Psittacosis is a zoonotic infection caused by Chlamydia psittaci. Most patients present with acute respiratory symptoms and systemic illness. When C. psittaci infects pregnant women, it causes severe clinical manifestations called gestational psittacosis. Here we report a case of gestational psittacosis. Our patient lacked respiratory symptoms, and pathological postmortem examinations revealed severe placentitis. Both DNA and immunohistochemical analyses were positive for C. psittaci from formalin‐fixed paraffin‐embedded tissues. The chlamydial DNA in the placenta was about 100 times more abundant than that in the lungs; therefore, the placenta rather than the lungs was the probable target of the C. psittaci infection during this pregnancy. We could not identify the source of infection. Gestational psittacosis should be considered in the differential diagnosis for fever of unknown origin during pregnancy, even in cases lacking respiratory symptoms. [ABSTRACT FROM AUTHOR]

Background: To minimize the risk of disease transmission in cornea transplantation, donor screening for blood-derived viral infections is mandatory. Ideally, pre-mortem blood samples are used, but based on availability, cadaveric blood samples of cornea donors may also be used. However, serological and nucleic acid amplification tests (NATs) need to be validated for the use of cadaveric specimens. Methods: Hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV), human T-lymphotropic virus (HTLV) 1/2, and Treponema pallidum (syphilis)-specific serological and/or NAT assays were validated on different platforms (Abbott Alinity i, Alinity m, Roche Cobas 6800, and Roche Cobas AmpliPrep/Cobas TaqMan (CAP/CTM)) using (un)spiked paired pre- and post-mortem cornea donor blood samples from the same individual (up to 23.83 h after death) of 28 individuals in accordance with the specifications of the German Federal Institute for Vaccines and Biomedicines (Paul-Ehrlich-Institut [PEI]). In addition, routinely HBV-, HCV- and HIV-PCR-negative tested post-mortem blood samples of 24 individuals were used to assess NAT specificity. Results: For the majority of serological parameters on the Abbott Alinity i (HBsAg, anti-HBc, anti-HBs, anti-HCV, anti-HIV, anti-HTLV 1/2, and anti-Treponema pallidum), ratios of generated test results of (un)spiked paired pre- and post-mortem blood samples differed ≤25%, with an agreement of qualitative pre- and post-mortem test results ranging from 91.2 to 100%. For NAT parameters (HBV, HCV, and HIV) on the Cobas 6800, Alinity m, and CAP/CTM, no significant deviation in virus concentrations (factor >5) of spiked pre- and post-mortem blood samples could be observed. Ct-values of corresponding internal controls did also not differ significantly (>1.5 Ct-values). In addition, no false-positive test results were generated when specificity was assessed. Conclusion: Overall, fluctuations of test results for serological and NAT parameters in pre- and post-mortem blood samples examined in this study, were only limited and within the range of what is also observed when routinely testing fresh patient specimens. We conclude that all examined assays are eligible for the screening of blood samples taken up to about 24 h after the occurrence of death. [ABSTRACT FROM AUTHOR]

Objectives: It has been generally believed that women who exposed to Toxoplasma gondii before pregnancy and have anti‐T. gondii IgG antibody are immunized and their newborns will be protected from congenital infection. This study is aimed to investigate the role of T. gondii infection in spontaneous abortion through serological and molecular methods in southern Iran. Study design: Blood samples were taken from 50 spontaneously aborted mothers and anti‐T. gondii antibodies were assessed using conventional enzyme‐linked immunosorbent assay (ELISA) and avidity ELISA methods. The placenta and blood samples of aborted women were used for detection of the parasite's DNA by polymerase chain reaction (PCR) method targeting the RE gene. The parasite genotypes were determined by PCR‐restriction fragment length polymorphism (RFLP) method using SAG3 and GRA6 genes. Results: IgG antibody was detected in 28% (14/50) of mothers, but all samples were negative for IgM antibody. In the avidity ELISA test, 26% (13/50) of the samples had a high avidity index, suggesting chronic infection, while a low avidity index was detected in one case (2%), which suggests acute infection. The parasite's DNA was detected in 18% (9/50) and 14% (7/50) of blood and placenta samples, respectively. All DNA positive samples were IgG positive. All isolates were belonged to the T. gondii type III genotype. Conclusion: The results suggest that T. gondii seropositive women are not protected from congenital transmission. However, the results should be interpreted cautiously until further studies will be confirmed these results. [ABSTRACT FROM AUTHOR]

Background Syphilis diagnosis may be challenging, especially in the asymptomatic and early clinical stages. We evaluated the presence of Treponema pallidum DNA (TP-DNA) in various sample types to elucidate transmissibility during various syphilis stages. Methods The study was conducted at the Amsterdam Centre for Sexual Health. We included adult men who have sex with men (MSM), who were suspected of having syphilis. The 2020 European guidelines definitions were followed for the diagnosis and staging of syphilis. Using a polymerase chain reaction (PCR) targeting the polA gene of Treponema pallidum (TP-PCR), we tested the following study samples on TP-DNA: peripheral blood, oropharyngeal swab, ano-rectal swab, and urine. Results From November 2018 to December 2019 we included 293 MSM. Seventy clients had primary syphilis, 73 secondary syphilis, 86 early latent syphilis, 14 late latent syphilis, 23 treated syphilis, and 27 had no syphilis. TP-DNA was detected in at least 1 study sample in 35/70 clients with primary syphilis (2/70 peripheral blood, 7/70 oropharynx, 13/70 ano-rectum, and 24/70 urine); in 62/73 clients with secondary syphilis (15/73 peripheral blood, 47/73 oropharynx, 37/73 ano-rectum, and 26/73 urine); and in 29/86 clients with early latent syphilis (5/86 peripheral blood, 21/86 oropharynx, 11/86 ano-rectum, and 6/86 urine). TP-DNA was not detected in clients with late latent syphilis or treated syphilis, nor in clients without syphilis. Conclusions TP-DNA was frequently detected in various sample types in the absence of lesions. This is in line with the high transmission rate of syphilis and opens diagnostic opportunities for early presymptomatic syphilis stages. [ABSTRACT FROM AUTHOR]

Cases of monkeypox, a double-stranded DNA virus that is closely related to smallpox, have recently increased in non-endemic countries, prompting fears of a new health emergency. Tens of thousands of cases have now been reported globally, with the majority of locations not having historically reported monkeypox. Here we review the epidemiology, transmission, diagnosis, management and prevention of monkeypox. [ABSTRACT FROM AUTHOR]