Quantitation of group A rotavirus by real-time reverse-transcription-polymerase chain reaction: Correlation with clinical severity in children in south India

The epidemiology and pathogenesis of rota-viruses are not completely understood, although recent developments in polymerase chain reaction (PCR) techniques now make it possible to quantify the viral load during an infective episode and investigate its relevance to clinical features of the disease. We studied rotavirus-positive stool samples collected from 10 children without symptoms of gastroenteritis and from 81 children with acute gastroenteritis and in whom the clinical severity of disease was recorded. A semi-quantitative real-time reverse-transcription (RT)-PCR was used to estimate the rotavirus load and to assess its correlation with the Vesikari score for severity of diarrhoea. There was a significant negative correlation (r=-0.80, P<0.001) between severity and the PCR cycle at which the PCR amplicons were detectable (crossing point) on the assay, indicating that children with more severe diarrhoea excrete more virus than children with less severe disease. (C) 2004Wiley-Liss,Inc.