Isolation and genotypic characterization of rotavirus

Two out of 12 fecal samples from children were found to be positive for rotavirus. Both the strains of rotaviruses were successfully isolated on MA 104 cell line where characteristic CPE was observed after 3 passages. The presence of virus was confirmed by SDS-PAGE and RT-PCR. Genotypic characterization was carried out by amplification of VP7 and VP4 genes followed by 2 separate multiplex PCR for G genotyping and another 2 for P genotyping using genotype specific primers. These isolates were characterized as G1P[6] and G11P[8] by RT-PCR and confirmed by nucleotide sequencing. Epidemiological investigation in the field need to be performed continuously at molecular level to ascertain whether G11P[8] specificity will predominate in human, and also to know the nucleotide exchange between porcine and human strains of rotavirus. Such type of emergence of noval strains may lead to change or update of exhisting anti-rotaviral vaccine component strains. Therefore, proper molecular epidemiological investigation is required not only in humans but also in animals.