Rotavirus subgroup characterisation by restriction endonuclease digestion of a cDNA fragment of the VP6 gene

Abstract

Human rotavirus strains characterised with subgroup-specific monoclonal antibodies were amplified using a VP6-specific RT-PCR and amplicons of 379 bp size (nt 747-1126) were analysed by restriction fragment length polymorphism (RFLP) using the restriction endonuclease AciI. The restriction patterns were compared to the subgroups determined serologically and to VP6 genogroups determined through sequence analysis. RFLP discriminated successfully between VP6 genogroups I and II. None of the strains characterised serologically as either subgroup nonI,nonII or subgroup I+II were distinguishable from subgroup II on the basis of their restriction pattern or sequence analysis. The results obtained by RFLP correlated well with the VP6 genogrouping results, providing a more reliable method for subgrouping of rotaviruses than ELISAs using subgroup-specific antibodies.