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

与 Sapporo 1982 株抗原相关的人杯状病毒 RNA 聚合酶区域的核苷酸序列多样性。

基本信息

批准号：
07670881
负责人：
NAKATA Shuji
金额：
$ 1.47万
依托单位：
Sapporo Medical University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07670881/
关键词：
HuCV Sa 82 J Infantile gastroenteritis ELISA RDRP region RT-PCR Sequence analys Dot Blot Hybridization Human calicivirus

项目摘要

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.

根据RNA依赖性RNA聚合酶（RDRP）基因组分析，人类杯状病毒（HuCV）至少可分为3个基因组：基因组I为诺瓦克病毒（NV），基因组II为雪山病毒（SMV），基因组III为HuCV/札幌/82/Japan（HuCV/Sa/82/J）。3个基因组间RDRP区序列同源性低于70%，未发现抗原相关性。通过RT-PCR扩增1979年至1990年间从札幌儿童中收集的HuCV/Sa/82/J相关菌株的RDRP区域并进行测序。PCR产物的核苷酸和氨基酸序列显示上述样品和HuCV/Sa/82/J原型之间的高度一致性。这些数据表明HuCV/Sa/82/J相关病毒之间的抗原性和RDRP区域的序列相似性之间存在相关性。这种病毒已经在札幌流行了至少10年。在美国、英国和沙特阿拉伯检测到的这些毒株也显示出与病毒原型在RDRP区域的高度一致性。用来自HuCV/Sa/82/J的RDRP区域的cDNA探针建立了检测HuCV/Sa/82/J的斑点杂交法。该方法对HuCV/Sa/82/J和相关病毒具有特异性，在高严谨性条件下，NV cDNA和猫杯状病毒RNA全基因组与HuCV/Sa/82/斑点杂交法检测粪便标本中病毒的阳性率（21%）高于ELISA法（10%）。该方法具有特异性强、操作简便、试剂不受限制等优点，可用于HuCV/Sa/82/J及其相关毒株的流行病学和分子生物学研究。