Characterization of VP7 and VP4 genes of rotavirus strains: 1990-1994 and 2000-2002.

A total of 90 rotavirus-positive faecal specimens collected from patients hospitalized with diarrhoea in 1990-1994 (n=77) and 2000-2002 (n=13) were investigated for VP7 and VP4 genotypes. The specimens included 21 typable and 69 non-reactive or multireactive rotavirus strains as monitored by monoclonal antibody-based serotyping ELISA. Genotyping was carried out by multiplex PCR/sequencing using primers specific for both VP7 and VP4 genes. The contribution of common genotypes (G1P[8], G2P[4], G3P[8], G4P[8]) in causing rotavirus diarrhoea was 79.2% and 92.3% in the years 1990-1994 and 2000-2002, respectively, while G9P[8] infections were detected at lower levels (1.3% and 7.7%) at both time-points. There was a predominance of G1P[8] in 1990-1994 and of G2P[4] in 2000-2002. The detection of unusual rotavirus strains (G1P[6], G1P[4], G1P[19], G2P[8], G3P[4], G4P[6]) in 19.5% of the patients indicated a significant contribution of reassortants in causing diarrhoea in western India.

Culture adaptation of serotype G6 human rotavirus strains from hospitalized diarrhea patients in India.

During serotyping of fecal specimens positive for rotavirus from hospitalized diarrhea patients in Pune, India, about 10% showed multireactivity in enzyme immunoassay with monoclonal antibodies specific for serotypes G1-4, 6, 8, and 10. In order to characterize some of these, three fecal specimens from children and one from adult were culture adapted. All the isolates showed long RNA pattern, but three out of four isolates belonged to subgroup I and II and one, to subgroup I. The isolates were confirmed as G6 by neutralization assay and polymerase chain reaction. Nucleotide sequences of cDNA derived from the gene encoding the outer capsid protein, VP7 of two strains indicated >94% identity with G6, the serotype, generally associated with cattle. The isolates were more close to G6 RF strain, which is a bovine rotavirus, reported from France. This is a first report of isolation of bovine serotype, G6 from children as well as adults from India.

Group B rotaviruses similar to strain CAL-1, have been circulating in Western India since 1993.

Generally, group A rotaviruses are the most common cause of paediatric diarrhoea. However, group B rotavirus, adult diarrhoea rotavirus (ADRV), was found to be involved in epidemics of severe gastroenteritis in several areas of China during 1982-1983 and had resulted in more than one million cases among adults as well as older children. Human group B rotavirus has been rarely reported outside China, but has been detected first from five adults with diarrhoea in Kolkata, India during 1997-1998 (strain CAL-1). During epidemiological studies at the National Institute of Virology (NIV) on hospitalized diarrhoea patients at Pune, India, faecal specimens from patients of >5 years age, which were negative for group A rotavirus by ELISA were tested by polyacrylamide gel electrophoresis (PAGE). We detected rotavirus RNA migration patterns similar to that of group B rotavirus in three faecal specimens from adults, two from the specimens collected in 1993 and one in 1998 from sporadic diarrhoea cases. RT-PCR was carried out using primers derived from gene 8 which codes for the NS2 protein, followed by nested PCR, which confirmed the presence of group B rotavirus in all three specimens. The sequences of the PCR products of NIV specimens were compared with that of CAL-1, ADRV and IDIR (infectious diarrhoea of infant rat) belonging to group B rotaviruses. The sequence analysis of the PCR products showed the highest identity with CAL-1, which was reported from Kolkata, India during 1997--1998. The finding suggests that human group B rotaviruses have been circulating in Pune. India, since 1993. This emerging virus may lead to more severe disease among adults in India. There is a need for surveillance of group B rotavirus infections, especially in adult diarrhoea cases and seroepidemiological studies on group B rotavirus are required among humans and animals of Western Maharashtra, India.