Nucleotide sequence diversity in RNA polymerase region of human calicivirus antigenically related to Sapporo 1982 strain.

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Besed on genome analysis of the RNA dependent RNA polymerase (RDRP) region, it has been proposed that human calicivirus (HuCV) can be classified into at least three genogroups : genogroup I is represented by Norwalk virus (NV), genogroup II by Snow Mountain virus (SMV) and genogroup III by HuCV/Sapporo/82/Japan (HuCV/Sa/82/J) virus. Among three genogroups, the sequnece homology in RDRP region is less than 70% and antigenic relatedness has not been demonstrated. The RDRP region of HuCV/Sa/82/J-related strains collected from children in Sapporo between 1979 and 1990 was amplified by RT-PCR and sequenced. Nucleotide and amino acid sequences of the PCR products showed a high degree of identity among above samples and prototype of HuCV/Sa/82/J.These data indicate a correlation between the antigenicity and sequence similarity of the RDRP region among HuCV/Sa/82/J-related viruses. This virus has been circulating in Sapporo for at least 10 years. These strains detected in USA,UK and Saudi Arabia also showed a high degree of identity in the RDRP region to prototype of virus.A dot blot hybridization assay with a cDNA probe derived from the RDRP region of HuCV/Sa/82/J was developed for detection of HuCv/Sa/82/J.This assay was specific for HuCV/Sa/82/J and related viruses, and the sensitivity was about 10^5 physical particles or 10pg of cDNA.The entire genome of NV cDNA and feline calicivirus RNA did not hybridize under high stringent conditions of stringency with the HuCV/Sa/82/J cDNA.A higher positive rate for virus detection in stool samples was obtained with the dot blot assay (21%) than ELISA (10%). The dot blot assay is specific, easy to perform and advantageous with unlimited supply of reagents, and should be useful for epidemiological and molecular biological studies of HuCV/Sa/82/J and related strains.