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The RNA was extracted from faecal suspension using a standard phenol�Cchloroform extraction E-64 method and ethanol precipitation. The RNA was electrophoresed in 10% polyacrylamide gel (2?mm thick) with a discontinuous buffer system, for 16?h at 20?mA, and visualized by silver staining. G and P typing of rotavirus-positive samples was carried out by nested and multiplex PCR using consensus and type-specific primers as described by Gouvea et?al. 1990 [10] and Gentsch et?al. 1992 [11]. Briefly, genotyping was undertaken by RT-PCR using primers Con2 and Con3 for gene 4 (VP4) and Beg9 and End9 for gene 9 (VP7). The cDNA was then used for P type-specific nested multiplex PCR using primers for P1A[8], IT-1; P1B[4], 2T-1; 2A[6], 3T-1; P3[9], 4T-1; P4[10], 5T-1 and for G type-specific nested multiplex PCR using primers for G1, aBT1; G2, aCT2; G3, aET3; G4, aDT4; G8, aAT8; G9, aFT9; and the common primer RVG9. For sample size calculation, we assumed 20% positivity for rotavirus in patients with acute watery diarrhoea, as reported in a previous study from India [12]. To detect this with a precision of 5% and �� level of 95%, a minimum sample size of 246 patients was required. We continued recruitment for a longer period to obtain larger numbers of rotavirus-positive cases of acute watery diarrhoea, in order to study their genomic diversity. Data were entered twice into customized software and analysed using SPSS 13.0 statistical software (SPSS Inc., Chicago, IL, USA). click here Univariate analysis was carried out for base-line variables. Z-scores for weight-for-height, R428 height-for-age and weight-for-age were calculated by using the least mean square method in an epidemiology information program EpiInfo 3.5 (CDC, Atlanta, GA, USA [13]). Univariate association of rotavirus positivity in patients with diarrhoea was assessed by calculating OR along with 95% CI and p-value. A p-value of