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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.

Objectives A retrospective study was undertaken to investigate the circulating dengue virus (DENV) serotypes and genotypes in India in 2018. Methods In total, 4963 samples referred to virus research diagnostic laboratories (n=21), the Indian Council of Medical Research-National Institute of Virology (ICMR-NIV) and ICMR-NIV field units (n=2) for diagnosis of dengue in 2018 were tested using a real-time reverse transcription polymerase chain reaction assay for the presence of DENV serotypes. Representative samples were sequenced for the envelope (E) gene. Results Regional diversity was observed with regard to the dominant circulating serotypes. DENV-2 was found to be the most common serotype in many states. Thrombocytopenia, petechiae and malaise were associated with DENV-2 infection. Phylogenetic analyses of DENV E gene sequences revealed the circulation of genotypes I and V of DENV-1, two lineages of DENV-2 genotype IV, DENV-3 genotype III and DENV-4 genotype I. Conclusions This study found regional differences in the prevalence of circulating DENV serotypes in India, and provides baseline data for continuous molecular surveillance. Molecular surveillance may have implications for predicting large-scale outbreaks of dengue if regional shifts in the predominantly circulating serotypes and genotypes are detected during the early phase of the dengue season.

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 & objectives: Public health and diagnostic laboratories are facing huge sample loads for COVID-19 diagnosis by real-time reverse transcription-polymerase chain reaction (RT-PCR). High sensitivity of optimized real-time RT-PCR assays makes pooled testing a potentially efficient strategy for resource utilization when positivity rates for particular regions or groups of individuals are low. We report here a comparative analysis of pooled testing for 5- and 10-sample pools by real-time RT-PCR across 10 COVID-19 testing laboratories in India. Methods: Ten virus research and diagnostic laboratories (VRDLs) testing for COVID-19 by real-time RT-PCR participated in this evaluation. At each laboratory, 100 nasopharyngeal swab samples including 10 positive samples were used to create 5- and 10-sample pools with one positive sample in each pool. RNA extraction and real-time RT-PCR for SARS-CoV-2-specific E gene target were performed for individual positive samples as well as pooled samples. Concordance between individual sample testing and testing in the 5- or 10-sample pools was calculated, and the variation across sites and by sample cycle threshold (Ct) values was analyzed. Results: A total of 110 each of 5- and 10-sample pools were evaluated. Concordance between the 5-sample pool and individual sample testing was 100 per cent in the Ct value ≤30 cycles and 95.5 per cent for Ct values ≤33 cycles. Overall concordance between the 5-sample pooled and individual sample testing was 88 per cent while that between 10-sample pool and individual sample testing was 66 per cent. Although the concordance rates for both the 5- and 10-sample pooled testing varied across laboratories, yet for samples with Ct values ≤33 cycles, the concordance was ≥90 per cent across all laboratories for the 5-sample pools. Interpretation & conclusions: Results from this multi-site assessment suggest that pooling five samples for SARS-CoV-2 detection by real-time RT-PCR may be an acceptable strategy without much loss of sensitivity even for low viral loads, while with 10-sample pools, there may be considerably higher numbers of false negatives. However, testing laboratories should perform validations with the specific RNA extraction and RT-PCR kits in use at their centres before initiating pooled testing.

The number of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) cases is increasing in India. This study looks upon the geographic distribution of the virus clades and variants circulating in different parts of India between January and August 2020. The NPS/OPS from representative positive cases from different states and union territories in India were collected every month through the VRDLs in the country and analyzed using next-generation sequencing. Epidemiological analysis of the 689 SARS-CoV-2 clinical samples revealed GH and GR to be the predominant clades circulating in different states in India. The northern part of India largely reported the ‘GH’ clade, whereas the southern part reported the ‘GR’, with a few exceptions. These sequences also revealed the presence of single independent mutations—E484Q and N440K—from Maharashtra (first observed in March 2020) and Southern Indian States (first observed in May 2020), respectively. Furthermore, this study indicates that the SARS-CoV-2 variant (VOC, VUI, variant of high consequence and double mutant) was not observed during the early phase of virus transmission (January–August). This increased number of variations observed within a short timeframe across the globe suggests virus evolution, which can be a step towards enhanced host adaptation.

Introduction: Rapid antigen testing can help in early diagnosis of Influenza in resource limited settings in patients presenting with Influenza Like Illness (ILI). Aim: To evaluate the usefulness of Rapid Influenza Direct antigen Testing (RIDT) in comparison to Real Time Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) for diagnosis of Influenza virus infection in clinically suspected patients with ILI in a tertiary care hospital. Material and Methods: A prospective observational independent study was done to evaluate RIDT for Influenza virus infection, considering RT-PCR as gold standard. Throat swabs and nasal swabs from patients meeting Category 3 definition of ILI by Ministry Of Health and Family Welfare (MOHFW) were collected from July 20th, 2019 to October 20th, 2019 from hospitals in and around the study area. Onsite Rapid test (Biogenix, China) was used as per manufactures instructions for rapid detection of Influenza antigen. RNA extraction was done using Qiagen kit followed by RT-PCR using primer, probes recommended by National Institute of Virology (NIV, Pune) as per their protocols. Data was recorded on a predesigned proforma and managed using Microsoft Excel 2018 (Microsoft Corp, Redmond, WA). The Statistical software MedCalc version 9.1 for Windows was used for data analysis. Results: The overall sensitivity of the RIDT was 37.9% and specificity was 94.4%. The Positive Predictive Value (PPV) and Negative Predictive Value (NPV) were 73.3% and 78.8%, respectively. Kappa was calculated to be 0.37 that suggested a fair agreement between RIDT and RT-PCR. When the Cycle threshold (Ct) values were compared to the results of RIDT, there were more false positives at low Ct value and more false negatives at high Ct values. Conclusion: The RIDT is not useful in screening during an outbreak in acute care settings. The negative result of RIDT should be followed up with RT-PCR. With a strong clinical suspicion, it is better to start antivirals than wait for laboratory report in severely ill patients with ILI.

A major limitation to providing oxygen therapy by high flow nasal oxygen (HFNO) delivery devices is its availability and therefore as an alternative many clinicians use a standard non rebreathing face mask (NRBM) in order to oxygenate their patients where low-flow nasal oxygen or simple facemask oxygen is not providing adequate respiratory support to achieve the target peripheral oxygen saturation (SpOIn this prospective open labelled study, 122 COVID-19 patients presenting with acute hypoxaemic respiratory failure (AHRF) were randomised to receive either HFNO or NRBM to achieve the target SpOThe device failure rate was significantly higher in HFNO group (39% versus 21%, P = 0.030). Oxygen support with NRBM resulted in a reduced all mortality rate over HFNO (26.2% versus 45%) but the mortality rate after treatment failure in either group (HFNO or NRBM) remained high (91% versus 92%).Oxygen support with NRBM results in both reduced device failure rate and higher survival among patients of COVID-19 with AHRF.

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: The present study was designed to know the trends of acute phase dengue in the Rayalaseema region of Andhra Pradesh state. Methods: This was a retrospective study conducted at Sri Venkateswara Institute of Medical Sciences, a tertiary care teaching hospital in Tirupati, Andhra Pradesh. Samples received for dengue NS1 testing from January 2011 to December 2015 were analysed. Results: During the study period, 6182 serum samples were received from cases clinically suspected to have dengue fever. Of these 690 (11.2%) tested positive. Positivity was significantly higher among females compare to males (12.1% vs 10.4%; P = 0.037). Children (19.2%) were more affected than adults (12.2%) and elderly (3.6%) (P < 0.001). Conclusion: Acute dengue infections were high in children compared to other age-groups with preponderance during post-monsoon and monsoon periods.

Introduction: Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) are serious public health problems worldwide with estimated 350 million chronically infected people. Transmission of infection occurs through blood transfusion, needles, body fluids and sexual intercourse. To know the magnitude of transmission of any disease in a community and for its control and prevention; trend and study of its prevalence is most important. Aim: To analyse the overall trend of HBV and HCV infection and to estimate the seroprevalence in a tertiary care hospital of Southern India. Materials and Methods: This was a retrospective study conducted at the Department of Microbiology, Sri Venkateswara Institute of Medical Sciences (SVIMS), Tirupati, Andhra Pradesh, India. The data of 77,158 and 58,024 samples for the test of Hepatitis B surface antigen (HBsAg) and test for anti-hepatitis C antibody respectively, for a period of three years four months (from September 2013 to December 2016) was retrieved from Departmental registers and Hospital Information System. Statistical software IBM SPSS, Version 20.0 (IBM SPSS Statistic, Somers NY, USA) was used for statistical analysis. Results: A total of 77,158 samples were tested for HBsAg whereas, 58,024 were tested for the presence of anti-HCV antibody. Seroprevalence of HBV was 2.27% and for HCV, it was 0.73%. Conclusion: HBV infection is showing a decreasing trend and the HCV infection, a fluctuating trend. Attempt should be made to reduce the incidence of HBV and HCV by simple preventive measures like public education, screening of blood and blood products, increasing public awareness about importance of vaccination.

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.

Background: Respiratory tract infections are a common occurrence after open heart surgery, leading to prolonged hospital stay, morbidity and costing the hospital exchequer. This study was conducted to study the utility of postextubation endotracheal tube [ET] tip culture in providing an early-basis for starting evidence-based antibiotic therapy. Methods: We retrospectively reviewed the case records of 172 patients who had undergone heart surgery at our tertiary care teaching hospital for occurrence of post-operative infections from clinical and microbiological data. Results: Bacterial culture was positive in 67(39%) patients. Endotracheal tube patients, grew organisms that were sensitive to empirical antibiotic therapy, in 39 (58%) patients. In 28 (42%) patients organisms resistant to empirical therapy were grown; only 14 of the 28 patients (50%) were symptomatic. Staphylococcus was the most common organism isolated followed by Acinetobacter and Enterobacter. Pseudomonas aeruginosa isolates were sensitive to all the antibiotics tested. Resistance was high among Acinetobacter sp. All the Gram-negative bacteria were sensitive to tigecycline. Resistance for beta-lactam antibiotics ranged from 35%-66% with imipenem being the most effective antibiotic. Conclusion: Our observations provide useful information regarding the microbiology of respiratory infection occurring in post-operative patients who had undergone open-heart surgery. Post-extubation endotracheal tip culture appears to be a useful tool for reliable and accurate diagnosis and treatment of these infections.

Fungal infections are a common cause of morbidity and mortality among patients on continuous ambulatory peritoneal dialysis (CAPD). We present the clinical manifestations, and management of five CAPD patients with fungal peritonitis seen during the last seven years. Microscopy and culture of the peritoneal fluid tested positive for fungus in all patients. Filamentous fungi were identified as the causative fungi among four patients and Candida nonalbicans was the cause in one patient. Catheter was removed and appropriate antifungal therapy was given to all the patients. Four patients recovered and one died due to ventilator associated infection. Previously treated bacterial infections, old age more than 50 years, diabetes and hypertension were recognised as the risk factors leading to fungal infection among CAPD patients.

Rhytidhysteron is a saprophytic dematiaceous fungus which rarely infects humans. Though virtually all individuals are exposed, very few develop the disease. Only seven human cases are reported till date. The present case is the second case from South India. A 40-year-old immunocompetent female agricultural worker, presented with a swelling on the dorsum of the right hand. Fine needle aspiration cytology (FNAC) of the swelling revealed short, thick, branched septate fungal hyphae. The isolate was moderately slow growing; grayish white colonies were observed on Sabouraud's Dextrose Agar (SDA) slant. On further incubation, the colonies turned floccose, greyish black and the black pigment was observed on the reverse. Microscopy of lactophenol cotton blue tease mount showed thick, brown septate hyphae without any fruiting bodies. Molecular typing confirmed the isolates as Rhytidhysteron rufulum. Identification of all clinical isolates of nonsporulating fungi to genus level is necessary to identify rare fungi infecting humans.

Background & objectives: Scrub typhus is a vector-borne zoonotic infection caused by Orientiatsutsugamushi. Local epidemiology of the circulating serotypes of scrub typhus is not available from most parts of India. We conducted this study for the diagnosis of scrub typhus using IgM ELISA and to detect O. tsutsugamushi serotypes circulating in southern Andhra Pradesh, India. Methods: Samples were collected from patients clinically suspected to have scrub typhus and were subjected to IgM ELISA to measure IgM antibodies against O. tsutsugamushi. Nested polymerase chain reaction (PCR) was performed targeting strain-specific regions in ELISA-positive samples. Results: Of a total of 663 samples, 258 (38.91%) were found to be positive by IgM ELISA. Serotypes could be detected in 230 (34.69%) samples only. Only two serotypes, Karp and Kawasaki, were found in the serum samples, with the former being predominant. The dual infection of Karp and Kawasaki serotypes was found in seven patients. Other serotypes such as Gilliam, Kuroki and Kato were not detected in the samples. Interpretation & conclusion: The nested PCR products proved useful in presumptively identifying the endemic O. tsutsugamushi serotypes. The present study could be significant in understanding scrub typhus epidemiology in this region.

Background & objectives: Ventilator-associated pneumonia (VAP) is an important hospital-acquired infection with substantial mortality. Only a few studies are available from India addressing the microbiological aspects of VAP, which have been done with small study populations. This study was carried out in the intensive care units (ICUs) of a tertiary care hospital to assess the profile of pathogens and to determine the pattern of antimicrobial resistance. Methods: This was a retrospective study of clinically suspected cases of VAP. Over a three year period, a total of 247 cases in 2011, 297 in 2012 and 303 in 2013 admitted in ICUs on mechanical ventilation with clinical evidence of VAP were included in our study. The endotracheal aspirate samples from these suspected cases were subjected to quantitative culture technique, and colony count of ≥10[5] colony forming units/ml was considered significant. Antimicrobial susceptibility test for the isolates was done. Results: VAP rates of 44.1, 43.8 and 26.3 were seen in 2011, 2012 and 2013, respectively. In all the three years, non-fermentative Gram-negative bacilli were the predominant organisms, followed by Pseudomonas spp. and Klebsiella spp. Staphylococcus aureus exhibited a downwards trend in prevalence from 50.0 per cent in 2011 to 34.9 per cent in 2013. An increase in vancomycin-resistant enterococci was seen from 4.3 per cent in 2012 to 8.3 per cent in 2013, while methicillin resistance amongst the S. aureus crossed the 50 per cent mark in 2013. An increasing trend in resistance was shown by Pseudomonas spp. for piperacillin-tazobactam (PTZ), amikacin and imipenem (IPM). For the non-fermenters, resistance frequency remained very high except for IPM (33.1%) and polymyxin-B (2.4%). Interpretation & conclusions: Our findings show VAP as an important problem in the ICU setting. The incidence of multidrug-resistant pathogens was on the rise. The resistance pattern of these pathogens can help an institution to formulate effective antimicrobial policy. To have a comprehensive pan-India picture, multicentric studies are needed.

Background: Safe blood donation remains a challenge in resource limited countries. False positive serological tests lead to wastage of large number of blood units. Nucleic acid amplification test (NAAT) technology has greatly enhanced the accuracy in identification of transfusions transmitted infections. Methods: The present study was undertaken to study the seroprevalence of hepatitis C virus (HCV) and asses the concordance between seropositivity with the presence of HCV ribonucleic acid (RNA) and to know the distribution pattern of HCV genotypes in healthy blood donors. Results: Among the 9287 donors screened (88.3% males), 7153 (77%) were voluntary donors while 2134 (23%) were replacement donors. Among blood donors, 27 (0.3%) samples tested HCV seropositive. Among 27 anti-HCV positive samples only 11 (41%) were found positive for HCV RNA. Among the 5 samples subjected to sequencing, three were found to be genotype 1a while two were genotype 3a. Conclusions: Our observations suggest that implementing NAAT test for HCV screening will be helpful in minimizingfalse-positive test results in the Indian setting.

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.

Background & objectives: Scrub typhus is a vector-borne zoonotic infection caused by Orientia tsutsugamushi. Local epidemiology of the circulating serotypes of scrub typhus is not available from most parts of India. We conducted this study for the diagnosis of scrub typhus using IgM ELISA and to detect O. tsutsugamushi serotypes circulating in southern Andhra Pradesh, India. Methods: Samples were collected from patients clinically suspected to have scrub typhus and were subjected to IgM ELISA to measure IgM antibodies against O. tsutsugamushi. Nested polymerase chain reaction (PCR) was performed targeting strain-specific regions in ELISA-positive samples. Results: Of a total of 663 samples, 258 (38.91%) were found to be positive by IgM ELISA. Serotypes could be detected in 230 (34.69%) samples only. Only two serotypes, Karp and Kawasaki, were found in the serum samples, with the former being predominant. The dual infection of Karp and Kawasaki serotypes was found in seven patients. Other serotypes such as Gilliam, Kuroki and Kato were not detected in the samples. Interpretation & conclusion: The nested PCR products proved useful in presumptively identifying the endemic O. tsutsugamushi serotypes. The present study could be significant in understanding scrub typhus epidemiology in this region.

Background: Detection of antinuclear antibody (ANA) is used as one of the diagnostic criteria for autoimmune rheumatic diseases (ARD). Both indirect immunofluorescence (IIF) and enzyme linked immunosorbant assay (ELISA) methods are used for this purpose. However, there are lack of data comparing these two tests from India. Methods: We prospectively studed 294 patients clinically suspected to be having ARD between April 2012 and September 2013. They were tested for ANA by IIF and ELISA methods. Representative samples positive by both the tests were processed again by a line immunoassay test to detect the specific antinuclear antibodies. Considering the IIF results as the ‘gold standard’, the utility of ELISA for ANA detection was analyzed. Results: Of the 294 samples processed, 181 (61.5%) were from female patients. By IIF 30% of samples in males and 40.3% sample in females tested positive. We found ELISA to have a poor sensitivity (45.8%) but good specificity (99.5%). The positive predictive value for ELISA were 98% and negative predictive value 76.2% respectively. Forty four samples positive by both IIF and ELISA were tested by Western blot to detect individual autoantibodies. Of these, only 24 samples showed the presence of one or more bands, while the remaining 20 (45.4%) were negative by line immunoassay. In our study anti-nuclear ribonucleoprotein/Smith was the most common ANA detected. Conclusions: The poor sensitivity raises concerns regarding the practice of initial screening for ANA by ELISA.

Fungal peritonitis is an uncommon condition which is associated with high morbidity and mortality in patients on continuous ambulatory peritoneal dialysis (CAPD). It is associated with several complications and many of the patients who develop this condition are unable to resume CAPD treatment and have to shift to haemodialysis. Here we report the rare occurrence of fungal peritonitis due to Aspergillus niger in a patient on CAPD.

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.

Background: Infections are the main reason for hospitalization and the second common cause of death in chronic kidney disease (CKD) patients. Infections develop primarily as a consequence of deranged innate immunity. In this study, we assessed the efficiency of phagocytic function of polymorphonuclear (PMN) leukocytes in severe CKD patients. Aims and Objectives: The aims of the study were (i) to assess the phagocytic index and lytic index of PMN leukocytes (PMNLs) in Stages 4 and 5 CKD patients. (ii) To compare the above indices with healthy age and sex-matched controls. Materials and Methods: The study was carried out in 60 adults in the age group of 18–60 years of which 30 were CKD patients in Stages 4 and 5 taking conservative treatment (not on dialysis) and other 30 were age and sex-matched controls. After screening for inclusion and exclusion criteria, 5 ml of venous blood was collected and taken immediately for evaluation. Phagocytic index which is the number of neutrophils positive for ingested microbes per 100 neutrophils and lytic index which is the total number of microbes per 100 cells were calculated. These are indices of neutrophil functioning. Results: Statistically significant decrease in both phagocytic index and lytic index were found in Stages 4 and 5 CKD patients as compared to controls. Conclusion: In the present study, we were able to establish that phagocytic capacity of PMNLs is adversely affected in severe CKD patients. This gains importance in the light of immune dysfunction being considered as a major cause of premature deaths resulting from infections in severe CKD.

Background & objectives: Dengue virus infection is endemic in India with all the four serotypes of dengue virus in circulation. This study was aimed to determine the geographic distribution of the primary and secondary dengue cases in India. Methods: A multicentre cross-sectional study was conducted at Department of Health Research / Indian Council of Medical Research (DHR)/(ICMR) viral research and diagnostic laboratories (VRDLs) and selected ICMR institutes located in India. Only laboratory-confirmed dengue cases with date of onset of illness less than or equal to seven days were included between September and October 2017. Dengue NS1 antigen ELISA and anti-dengue IgM capture ELISA were used to diagnose dengue cases while anti-dengue IgG capture ELISA was used for identifying the secondary dengue cases. Results: Of the 1372 dengue cases, 897 (65%) were classified as primary dengue and 475 (35%) as secondary dengue cases. However, the proportion varied widely geographically, with Theni, Tamil Nadu; Tirupati, Andhra Pradesh and Udupi-Manipal, Karnataka reporting more than 65 per cent secondary dengue cases while Srinagar, Jammu and Kashmir reporting as low as 10 per cent of the same. The median age of primary dengue cases was 25 yr [interquartile range (IQR 17-35] while that of secondary dengue cases was 23 yr (IQR 13.5-34). Secondary dengue was around 50 per cent among the children belonging to the age group 6-10 yr while it ranged between 20-43 per cent among other age groups. Interpretation & conclusions: Our findings showed a wide geographical variation in the distribution of primary and secondary dengue cases in India. It would prove beneficial to include primary and secondary dengue differentiation protocol in the national dengue surveillance programme.