Search

Due to a wide range of Human Papillomavirus (HPV) types associated with genital cancers; HPV genotyping remains important for the introduction of an appropriate vaccine, disease diagnosis, follow-up and epidemiological surveys. Currently, available molecular genotyping assays are not only expensive but also requires dedicated and expensive equipment which is not feasible in the majority of low-and-middle-socioeconomic countries. The purpose of the study was to develop and evaluated a cost-effective nested-multiplex polymerase chain reaction (NM-PCR) assay for HPV genotyping.HPV-DNA containing plasmids and cervical scrapings from histologically confirmed cervical cancer cases were used to evaluate the NM-PCR. In the first round PCR, a set of consensus primers were used to amplify 38 mucosal HPV types. HPV Type-specific primers were used in the second-round polymerase chain reaction (PCR) to amplify 15 HPV types in three multiplex cocktails. The assay sensitivity was determined with the control panel containing one to 10The assay was able to amplify all HPV types and detected as few as 50GE per reaction. A total of 23 endo-cervical samples obtained from healthy, HPV negative subjects and 52 histologically confirmed cervical scrapings were processed for HPV genotyping by NM-PCR. HPV DNA was detected in all histologically confirmed samples. DNA sequencing results showed complete concordance with PCR results.The designed nested PCR based assay had good concordance with clinical histology and sequencing results and appears to be a promising tool for HPV genotyping especially in resource-constrained settings.

Human papillomavirus (HPV) causes genital and oropharyngeal cancers worldwide. There are significant gaps exist in the data on HPV genotype prevalence in this part of the country. HPV vaccination is one of the best preventive methods available currently. HPV genotyping plays an important role in the selection of appropriate vaccines and monitoring vaccine efficacy and coverage. The present study aimed to determine the HPV genotype prevalence and to estimate the potential impact of HPV vaccines on invasive cervical cancer.A total of 204 cervical biopsy samples collected from symptomatic women were subjected to an in-house designed and standardised nested multiplex PCR (NM-PCR) assay. The NM-PCR was designed to detect 38 Mucosal HPV types as a pooled result and genotyping of 15 HPV types. Further, the HPV genotype data was used to estimate the HPV vaccine bivalent, quadrivalent and nonavalent impact on the population using a mathematical formula.Out of 204 samples 188 were subjected to HPV-nested PCR. A total of 163 (86.7%) samples were positive for at least one HPV type. Multiple genotypes were identified in 30% of samples processed. HPV-16 (85.3%) was the most frequently detected genotype followed by HPV-18 (13.5%) and HPV-33 (11.0%). Other genotypes were observed less frequently. Based on the HPV prevalence observed in the study a mathematical model estimated the efficacy of bivalent, quadrivalent and nonavalent vaccines were 76.1%, 76.7%, and 91.1% (average) respectively.HPV-16 was the most prevalent (85%) genotype detected in this study. Multiple infections observed in 30% of samples were quite high as compared to the majority of national, and global reference (15-25.4%) data. The Mathematical model showed that a nonavalent vaccine would give better protection.

A large number of new molecular or virology laboratories have been established to increase the testing capacity for SARS-CoV-2. Due to heavy workload, there is delay in testing of samples. In order to avoid the negative effect of delayed testing on RTPCR results guidelines are issued from WHO and CDC to transport samples in cold chain. However, in pandemic situations the recommended guidelines for transport and storage conditions are often compromised. This study was conducted to evaluate the effect of sample storage conditions at different temperatures on the results of RT PCR test.Total 275 samples were included in this study, among these 126 samples tested positive and 149 samples tested negative. All samples were aliquoted into two and the aliquotes stored in duplicate at 4 ​°C and room temperature. All aliquots stored at both the temperatures were tested by RTPCR every 24 hours up to 5 days.Diagnostic accuracy decreased from day1 to day 5 at both the storage temperatures i,e 4 ​°C and room temperature in comparison to the initial day results. Positivity decreased on an average of 9.02% at 4 ​°C and at 9.27% at room temperature per day. Among total 126 positive samples on an average false negative and failure of internal control at 4 ​°C and room temperature was 8.86%, 8.22% and 3.64%, and 4.12%, respectively. All the samples with CT value ​ ​30 remained positive at both temperatures up to 5 days. Few samples with30 CT value showed variable results i.e. positive, negative or internal control failure from day 1 (2nd day after sample collection) onwards.There was no significant difference between RT PCR results of samples stored at 4 ​°C and room temperature up to 5 days of collection. However internal control failure was more in samples stored at room temperature. Therefore, samples received without cold chain also may be processed by RTPCR and should not be rejected.

Background: Dengue is one of the most important mosquito-borne viral diseases in the world. The emergence and spread of four dengue viruses (DENVs) (serotypes) represent a global pandemic. The four distinct serotypes are, namely, DENV-1, DENV-2, DENV-3 and DENV-4. Very few dengue serotyping studies have been reported from Andhra Pradesh. In this context, the present study focuses on the circulating serotypes of dengue in South-Eastern Andhra Pradesh. Methodology: Study was done at Sri Venkateswara Institute of Medical Sciences, a teaching hospital in Tirupati, Andhra Pradesh. Acute phase dengue serum samples were collected and tested for NS1 antigen and anti-human IgM antibodies by enzyme-linked immunosorbent assay (ELISA). NS1-positive samples were further serotyped by reverse transcriptase real-time polymerase chain reaction (rRT-PCR). Results: A total of 398 serum samples were received from clinically suspected dengue fever cases. Of these, 150 (37.7%) samples were positive for NS1 and/or IgM ELISA. The 96 NS1 antigen-positive samples were further processed for serotyping, of which 36 were negative by rRT-PCR. DENV-2 (41%) was the predominant serotype, followed by DENV-4 (37%), DENV-3 (12%) and DENV-1 (10%) in descending order. Conclusion: This study reports the all four dengue serotypes' co-circulation. This is the first report from South Eastern Andhra Pradesh. Amongst four, DENV-2 was predominant followed by DENV-4. The information of predominant serotypes can guide in forecasting dengue outbreaks and improving control measures of vectors thus may be helpful in the prevention of outbreaks.

Background and Objectives: Influenza virus is a typical human pathogen causing serious respiratory illness resulting in significant mortality throughout the globe. Andhra Pradesh witnessed the first case of influenza A H1N1 in India from Hyderabad (now in Telangana) on May 16, 2009. In the recent past, Andhra Pradesh witnessed exponential increase in the number of confirmed cases of influenza infection. In this study, we present the salient features of the recent outbreak of influenza during 2017–2018 in the state of Andhra Pradesh, first of its kind after the division of the state. Materials and Methods: Clinically, suspected cases of influenza-like illness received in the Virus Research and Diagnostic Laboratory, Department of Microbiology, Sri Venkateswara Institute of Medical Sciences (SVIMS), Tirupati, from January 2017 to May 2018 were included in the study. The samples were tested for influenza A, influenza A (H1N1) pdm09, influenza A (H3N2), influenza B, influenza B/Yamagata and influenza B/Victoria. Results: A total of 1286 samples were received for testing. The positive samples were influenza A unsubtypable (109), influenza A (H1N1) pdm09 (356), influenza A (H3N2) (38) and influenza B (19; Victoria - 2, Yamagata - 17). There was no significant difference in positivity between genders with 260 (49.81%) females and 262 (50.19%) males being positive. Conclusion: The outbreak started in the late monsoon (January) of 2017 and had two peaks; one in summer months and another in winter months. Influenza B virus was reported from December 2017 to May 2018. Age groups ≤5 years and 6–18 years had higher positivity as compared to other age groups. Regular surveillance programmes are required for assessing the trends of influenza infections due to various subtypes and to plan timely and adequate steps for preventing the spread to larger vulnerable population.

Introduction: Scrub typhus is a rickettsial infection which is caused by Orientia tsutsugamushi and transmitted by the bite of the chigger of a mite. Delay in diagnosis can be fatal otherwise the treatment is simple, doxycycline being the drug of choice. Indirect immunoflurescence is considered gold standard but it is not used in India as it is costly and also not available. There is need for rapid, economic and simple test for the diagnosis of scrub typhus. This study was taken up to study the seroprevalence of scrub typhus in Andhra Pradesh and to compare two commonly used serological methods; rapid test and IgM ELISA. Materials and methods: This is a prospective study in which 100 serum samples from clinically suspected cases collected over a period of 3 months were processed for the detection of IgM antibodies for scrub typhus by ELISA and Rapid test. Samples were also tested for leptospirosis and dengue fever which the other common causes of fever prevalent in this region. Results: Total number of samples processed was 100 of which 52 were males and 48 females. Among the hundred samples 39 were seropositive. Positivity was higher in the age group of patients between 16 and 30 yrs of age. There was 97% correlation between ELISA and rapid method. Of the 100 samples only three samples positive by ELISA were negative by rapid method. Fever was the most common manifestation and there was no eschar and no mortality reported. Conclusion: Scrub typhus should be included in the differential diagnosis of fever of unknown origin along with dengue, malaria and leptospirosis which are the other common endemic infections in this part of the country.

Purpose: Coryneform or the non-diphtherial Corynebacterium species largely remains a neglected group with the traditional consideration of these organisms as contaminants. This concept, however, is slowly changing in the light of recent observations. This study has been done to find out the species distribution and antibiogram of various members of the clinically relevant Coryneform group, isolated from various clinical materials. Materials and Methods: One hundred and fourteen non-duplicate isolates of diphtheroids from various clinical isolates were selected for the study. The isolates were identified to the species level by using a battery of tests; and antimicrobial susceptibility was tested by using a combination of Clinical and Laboratory Standards Institute (CLSI) and the British Society for Antimicrobial Chemotherapy (BSAC) guidelines, in the absence of definitive CLSI guidelines. Results: Corynebacterium amycolatum was the predominant species (35.9%) in our series followed by the CDC Group G organisms (15.7%). Each of the remaining 19 species comprised of less than 10% of the isolates. More than half the total isolates were resistant to the penicillins, erythromycin, and clindamycin; while excellent activity (all the strains being susceptible) was shown by vancomycin, linezolid, and tigecycline. Chloramphenicol and tetracycline also had good activity in inhibiting more than 80% of the isolates. Multiply drug resistance was exhibited by all the species. Conclusion: This study was an attempt to establish the clinical significance of coryneform organisms. The high level of resistance shown by this group to some of the common antibacterial agents highlights the importance of processing these isolates in select conditions to guide the clinicians towards an appropriate therapy.

Fungal infections are being increasingly reported from immuno-compromised as well as immuno-competent patients. Transplant patients are on long term immunosuppressive therapy which makes them highly vulnerable to opportunistic fungal infections .These infections can be cutaneous or systemic. Several fungi have been reported to be the culprits such as Candida spp., Aspergillus spp., C. neoformans, P. carinii, and zygomycetes group of fungi. Cutaneous infections are most commonly caused by Pityriasis (tinea) versicolor, dermatophytes, and candida sp but these days the demtiaceous fungi are becoming more frequently reported .Here we report a case of post renal transplant cutaneous infection caused by dematiaceous fungus belonging to the order Pleosporales.