Molecular characterisation of Hepatitis E virus isolates from north India.

PURPOSE: Hepatitis E virus (HEV) is responsible for >50% of acute viral hepatitis (AVH) in developing countries. It has 4 major genotypes and various subtypes which vary in geographical distribution, clinical manifestations and epidemiological patterns. This study was conducted to characterise HEV isolates from north India to study the effect of host and viral factors on HEV infection. METHODS: Serum samples collected from 536 AVH patients admitted to Department of Medicine, King George's Medical University, Lucknow from July 2016 to June 2017 were screened for anti HEV IgM, anti HAV IgM, HBsAg and anti HCV antibodies using commercial ELISA kits. Samples either positive for anti HEV IgM antibodies (n â€‹= â€‹204) or negative for all 4 hepatotropic viruses (n â€‹= â€‹37) were enrolled and tested by real time PCR for HEV RNA. HEV RNA positive samples with high viral load were further subjected to nested PCR for amplification of capsid gene. Sequencing and phylogenetic analysis were performed. HEV strains isolated from this study were deposited to GenBank under accession numbers MG571274 to MG571283. RESULTS: Anti HEV IgM positivity was observed among 38% clinically suspected AVH cases. HEV RNA was detected in 31.8% seropositive HEV cases and additional 3 seronegative cases. Males outnumbered females and the most affected age group was of young adults. Maximum number of cases were seen during the months of June to September. Phylogenetic analysis showed that HEV strains in our study belonged to genotype 1a. Mortality in HEV infected pregnant females was 23.5% as compared to 2.4% in non-pregnant females. Adverse fetal outcome was recorded in 51% of HEV infected pregnancies. CONCLUSIONS: HEV genotype 1a is prevalent in our setting. HEV during pregnancy is associated with adverse maternal and fetal outcome.

Second Line Injectable Drug Resistance and Associated Genetic Mutations in Newly Diagnosed Cases of Multidrug-Resistant Tuberculosis.

Aim: To investigate the phenotypic and genotypic profile of multidrug-resistant (MDR) Mycobacterium tuberculosis (MTB) clinical isolates with reference to second-line injectable drugs (SLIDs). Methods: A total of 110 MTB isolates, recovered consecutively from confirmed MDR-TB patients between March and June 2016, were included in this study. Phenotypic drug susceptibility testing against SLIDs (Kanamycin, Amikacin, and Capreomycin) and Ofloxacin (OFX) was performed using the MGIT 960 system. For genotypic analysis, SLID/(s) resistant (n = 13) and susceptible isolates (n = 26) were subjected to PCR and DNA sequencing for rrs, eis (promoter region), and tlyA loci of MTB. Furthermore, the identified genetic mutations were analyzed with respect to its significance in detecting phenotypic resistance. Result: Among the 110 analyzed isolates, phenotypic resistance to OFX, SLIDs, and to both was 59.1%, 11.8%, and 10.0%, respectively. Out of a total 13 SLID/(s) resistant isolates, 10 had mutations (including two novel mutations) in one or more of the targeted genes. Only one SLID susceptible MTB isolate showed mutation in the targeted region. In SLID resistant isolates, most frequent mutation detected was C-12T under eis promoter region (46.1%). Conclusion: Mutations in rrs, eis, and tly A loci together are important in predicting SLID resistance in MTB isolates. Future molecular epidemiology studies are needed to have more insight into frequency and clinical relevance of novel mutations identified in this study.

Human Parvovirus 4: A harmless bystander or a pathogen of severe acute respiratory illness.

INTRODUCTION: Severe Acute Respiratory Infection (SARI) is an important cause of morbidity and mortality worldwide, caused by a large number of viral and bacterial agents. PARV4 is a recently identified virus detected in human blood and variety of tissues, but its disease association with SARI could not be established. OBJECTIVE: In the present case control study, we aim to investigate the association of PARV4 with SARI. METHODS: The Nasal and Throat swab (NS/TS) samples of 241 cases and 146 healthy controls were tested for most common respiratory viruses and PARV4 by real-time PCR. RESULTS: PARV4 was detected in 64(26.55%) SARI cases and only one healthy control (0.68%). PARV4 was the most common viral agent detected in SARI cases. A strong association of PARV4 is seen with severe respiratory illness. CONCLUSION: Detection of PARV4 in a significantly higher number of SARI cases, in comparison with controls, suggests association of PARV4 with SARI. PARV4 genotype 2 is the only circulating strain detected in our study.

Distribution of hepatitis C genotypes in Uttar Pradesh, India; rare genotype 4 detected.

AIM: The worldwide prevalence of hepatitis C virus infection (HCV) is nearly 150 to 170 million cases. The prevalence of HCV infection in India is estimated to be around 1%. In India HCV genotype (GT)3 is the predominant GT followed by GT1. Our study aims to establish the prevalent GTs/subtypes of HCV circulating in Uttar Pradesh, North India, as reported from a tertiary care hospital. METHODS: The study was a retrospective observational analysis of consecutive 404 HCV RNA positive cases referred to our hospital from September 2014 to April 2017, and was approved by an institutional ethics committee. Written informed consent was taken from each participant. Clinical and demographic details of these patients were recorded using predesigned questionnaires. All the laboratory testing was carried out on a stored serum sample of enrolled cases. Genotyping of all 404 strains was done by Sanger's sequencing of the core region. The phylogenetic analysis of 179 HCV strains with a high-quality sequencing data was performed. RESULTS: The distributions of prevalent GTs/subtypes as noted in the current study were ( n [%]): GT1a, 101 (25%); GT1b, 12 (2.9%); GT1c, 1 (0.25%); GT3a, 275 (68.07%); GT3b, 9 (2.2%); GT3g, 2 (0.49%); GT3i, 3 (0.74%); and GT4a, 1 (0.24%). HCV GTs GT2, GT5, and GT6 were not detected from our region. Sequence analysis showed high genotypic variability in HCV GT3. Phylogenetic analysis showed that HCV GT3 and GT1 circulating in our region were related to Indian strains reported earlier. CONCLUSIONS: HCV GTs 3a and 1a are the commonest circulating GTs in Uttar Pradesh, India.