Characterisation of rotavirus strains identified in adolescents and adults with acute gastroenteritis highlights circulation of non-typeable strains: 2008–2012

AbstractBackgroundGroup-A Rotavirus (RV) is the main causative agent of acute gastroenteritis in children < 5 years of age. Its role as a pathogen in adults needs to be monitored. The aim of this study was to characterise the group-A RV strains that cause infections of acute gastroenteritis in adolescents and adults and determine the temporal variations in the circulating strains during 2008–2012 in continuation of an earlier study conducted in 2004–2007, in Pune, India.MethodsA total of 371 stool samples were tested by RV antigen capture ELISA. VP4, VP6, VP7 and NSP4 genes of all of the RV strains detected in the study were analysed using reverse transcription PCR, multiplex PCR and sequencing.ResultsGroup-A RV was detected in 9.4% (35/371) of the stool samples examined in the study period. The frequency of detection of RV was found to decline from 18.0% (16/90) in 2008 to 3.8% (2/52) in 2012. Of the 6 strains typed for both VP7 and VP4 genes, G2P[4], G1P[8] and G9P[4] were detected in 3, 1 and 2 samples, respectively. Sequencing and phylogenetic analysis of the VP4, VP6, VP7 and NSP4 genes revealed an infrequently reported NSP4-E6 genotype and circulation of heterogenous [G2 (lineage IIC and IID), G9 (lineage 3), P[4] (lineage P[4]-5), P[8] (lineage P[8]-3), VP6 I1 / I2 and NSP4 E2] genotypes/lineages in the RV strains. Analysis of linkage within these genes showed concordance (G2-P[4]-I2-E2) and discordance (G9-P[4]-I2-E6), equally. The sequences of amplified VP6 (n = 20) and NSP4 (n = 2) genes from G and P nontypeable RV strains (80.0%, 28/35) were most homologous to human group-A RV strains.ConclusionThe study underscores the significant temporal variations in RV strains, identifies circulation of intergenogroup reassortants among adolescent and adult patients with acute gastroenteritis and emphasizes the need for continued surveillance and whole genome analysis of emerging rotavirus strains

Changing trends in circulating rotavirus strains in Pune, western India in 2009–2012: Emergence of a rare G9P[4] rotavirus strain

AbstractBackgroundA vast diversity in rotaviruses at inter- and intra-genotypic level underscores the need for monitoring of circulating rotavirus strains. The aim of this study was to update the data on rotavirus disease and strains for the period from January 2009 to December 2012 in Pune, western India which has been one of the sites of the Indian Rotavirus Strain Surveillance Network since November 2005.MethodsChildren aged <5 years admitted for acute gastroenteritis in three different hospitals from Pune city were included in the study. The stool specimens were collected and tested for rotavirus antigen by a commercial enzyme immunoassay. The rotavirus strains were genotyped by multiplex reverse transcription polymerase chain reaction.ResultsDuring the study period, we found 35.1% of 685 stool specimens contained rotavirus antigen. Frequency of rotavirus detection was greatest (58.5%) among children aged 7–12 months. The G1P[8] (31.4%), G2P[4] (20.2%) and G9P[8] (11.8%) strains were the most common types. We noted predominance of G1P[8] strains (39.6%-46.1%) in all the years of study except 2009 wherein G9P[8] strains scored highest level (15.3%). Subsequent to this, we identified G9P[8] strains at the second highest position in 2010, their sudden decline and rise in G9P[4] strains in 2011–2012. We detected G12 strains in combination with P[6] and P[8] at variable rates (0–10.2%) and highest level (27.1%) of mixed rotavirus infections in 2009 as compared to 2010–2012 (0–3.8%).ConclusionThe study highlights the huge burden of rotavirus disease and changing profile of circulating rotavirus strains displaying emergence of G9P[4] reassortant strains in Pune, western India and emphasizes the need to analyze the entire genomic constellation of rotavirus strains for better evaluation of the impact of rotavirus

Enteroviruses in Patients with Acute Encephalitis, Uttar Pradesh, India

An outbreak of viral encephalitis occurred in northern India in 2006. Attempts to identify an etiologic agent in cerebrospinal fluid by using reverse transcription–PCR showed positivity to enterovirus (EV) in 66 (21.6%) of 306 patients. Sequencing and phylogenetic analyses of PCR products from 59 (89.3%) of 66 specimens showed similarity with EV-89 and EV-76 sequences

Enteroviruses in Patients with Acute Encephalitis, Uttar Pradesh, India

An outbreak of viral encephalitis occurred in northern India in 2006. Attempts to identify an etiologic agent in cerebrospinal fluid by using reverse transcription–PCR showed positivity to enterovirus (EV) in 66 (21.6%) of 306 patients. Sequencing and phylogenetic analyses of PCR products from 59 (89.3%) of 66 specimens showed similarity with EV-89 and EV-76 sequences

Intragenotypic diversity in the VP4 encoding genes of rotavirus strains circulating in adolescent and adultcases of acute gastroenteritis in Pune, Western India: 1993 to 1996 and 2004 to 2007

International audienceGenetic variability of rotaviruses circulating in Pune, India at the two time points was determined by characterizing VP4 genes in 131 rotavirus strains detected in adolescent and adult cases of acute gastroenteritis. The multiplex RT-PCR classified the VP4 genes in 73 P[4] (43.2%), 69P[8] (40.8%) and 27 P[6] (16.0%) genotypes. Sequencing and phylogenetic analysis revealed increase in the prevalence of P[4]-5 and P[8]-2 and decline of P[8]-3 lineages in 2000s as compared to those identified in 1990s (92.8% Vs 100%, 4.2% Vs 33.3% and 93.7% Vs 66.7%, respectively). The P[4]-1 and P[8]-4 lineages circulated at low levels (7.1% / 2.1%) while presence of only P[6]-1 (100%) lineage was detected at both time. The strains with different VP4 genotype specificities displayed 0.2 to 2.3% amino acid divergence. A significant difference (P<0.01) in their association with common and nontypeable G strains was noted between the two time points studied. This is the first report to describe the intragenotypic diversity in the rotavirus VP4 genes from adolescent and adult patients of acute gastroenteritis from India. VP4 being one of the major protective antigens, monitoring the mutations in this protein would be crucial to understand the evolutionary changes in rotaviruses and devise more effective vaccine strategies in developing countries

Intragenotypic diversity in the VP4 encoding genes of rotavirus strains circulating in adolescent and adultcases of acute gastroenteritis in Pune, Western India: 1993 to 1996 and 2004 to 2007

International audienceGenetic variability of rotaviruses circulating in Pune, India at the two time points was determined by characterizing VP4 genes in 131 rotavirus strains detected in adolescent and adult cases of acute gastroenteritis. The multiplex RT-PCR classified the VP4 genes in 73 P[4] (43.2%), 69P[8] (40.8%) and 27 P[6] (16.0%) genotypes. Sequencing and phylogenetic analysis revealed increase in the prevalence of P[4]-5 and P[8]-2 and decline of P[8]-3 lineages in 2000s as compared to those identified in 1990s (92.8% Vs 100%, 4.2% Vs 33.3% and 93.7% Vs 66.7%, respectively). The P[4]-1 and P[8]-4 lineages circulated at low levels (7.1% / 2.1%) while presence of only P[6]-1 (100%) lineage was detected at both time. The strains with different VP4 genotype specificities displayed 0.2 to 2.3% amino acid divergence. A significant difference (P<0.01) in their association with common and nontypeable G strains was noted between the two time points studied. This is the first report to describe the intragenotypic diversity in the rotavirus VP4 genes from adolescent and adult patients of acute gastroenteritis from India. VP4 being one of the major protective antigens, monitoring the mutations in this protein would be crucial to understand the evolutionary changes in rotaviruses and devise more effective vaccine strategies in developing countries

Genetic analysis of rotavirus G2P[4] strains in Pune, Western India:circulation of a novel reassortant bearing E6 NSP4 genotype

AbstractIn India, G2P[4] strains are known to be the second most predominant group A rotaviruses causing acute gastroenteritisamong children. This study was performed to determine the diversity within VP7(G), VP4(P), VP6(I) and NSP4(E) genes of16 G2P[4] rotavirus strains detected in children hospitalized for acute gastroenteritis in Pune, Western India during 2009-2013. Fourteen strains showed G2-P[4]-I2-E2 and two strains showed G2-P[4]-I2-E6 genotype constellation. Phylogeneticanalysis showed their clustering into G2-IV-a3, P[4]-5bi/ii, I2-3ii and E2-4i/ii or E6 genotypes/lineages. These data revealinter- and/or intra-genotypic variations in a genogroup-2 constellation of G2P[4] rotavirus strains circulating in Pune, WesternIndia, providing evidence of a novel G2P[4] reassortant bearing a rare NSP4 genotype, E6 during 2009-2013

Evaluation of Urine as a Clinical Specimen for Diagnosis of Hepatitis A

The present study pertains to the evaluation of urine as a specimen for detection of anti-hepatitis A virus (anti-HAV) antibodies. Immunoglobulin M (IgM), IgG, and IgA capture enzyme-linked immunosorbent assays for hepatitis A were performed on paired serum and urine specimens collected from hepatitis A patients (n = 92), healthy individuals (n = 100), non-A hepatitis patients (n = 70), and patients with nonhepatic diseases (n = 64, including 37 renal disease patients). Hepatitis A patients seropositive for anti-HAV IgM showed 95.65% uropositivity. No false-positive reactions were observed in control groups. The uropositivity of anti-HAV IgM persisted during the convalescent phase of the disease. Anti-HAV IgG uropositivity correlated well with corresponding seropositivity in all groups (P > 0.05 for each). No significant difference between the proportions of serum and urine positivity for anti-HAV IgA was noted (P > 0.05 for each). Using seroreactivity as a “gold standard,” the sensitivity and specificity for anti-HAV IgM, anti-HAV IgG, and anti-HAV IgA tests with urine as a specimen were found to be 95.65 and 100%, 97.76 and 76.47%, and 92.23 and 88.18%, respectively. Urine appears to be comparable to serum for diagnosis of recent and past infection with hepatitis A

Direct costs of hospitalization for rotavirus gastroenteritis in different health facilities in India

Background &#38; objectives: Diarrhoeal disease is the fifth leading cause of all mortality globally. To this burden, rotavirus contributes over half a million deaths annually. This pilot study was conducted to determine the economic burden of diarrhoeal episodes on families from different geographical regions accessing medical facilities in India. Methods: Participants were enrolled from four study sites with eight reporting hospitals, categorized as non-profit and low cost, private and government facilities between November 2008 and February 2009. Questionnaires detailing healthcare utilization, medical and non-medical expenditure and lost income were completed by families of children &#60; 5 yr of age hospitalized for gastroenteritis. All available faecal samples were tested for rotavirus. Results: A total of 211 patients were enrolled. The mean total cost of a hospitalized diarrhoeal episode was ` 3633 (US$66.05) for all facilities, with a marked difference in direct costs between governmental and non-governmental facilities. Costs for rotavirus positive hospitalizations were slightly lower, at ` 2956 (US$ 53.75). The median cost of a diarrhoeal episode based on annual household expenditure was 6.4 per cent for all-cause diarrhoea and 7.6 per cent for rotavirus diarrhoea. Of the 124 samples collected, 66 (53%) were positive for rotavirus. Interpretation &#38; conclusions: Data on direct costs alone from multiple facilities show that diarrhoeal disease constitutes a large economic burden on Indian families. Affordable, effective vaccines would greatly reduce the economic burden of severe gastroenteritis on patients, families and the government