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Use of genomic probes to detect hepatitis A virus and enterovirus RNAs in wild shellfish and relationship of viral contamination to bacterial contamination.

机译:使用基因组探针检测野生贝类中的甲型肝炎病毒和肠道病毒RNA以及病毒污染与细菌污染的关系。

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摘要

Genomic probes were used to investigate hepatitis A virus (HAV) and enterovirus RNAs in two types of shellfish from natural beds (Atlantic coast, France). After elution concentration, nucleic acid extracted by proteinase K and purified by phenol-chloroform and ethanol precipitation was assayed by dot blot hybridization. The probes used were a specific HAV probe corresponding to the 3' end (3D polymerase coding region) and an enterovirus probe corresponding to the 5' noncoding region. The method was first tested under experimental conditions by using virus-spiked shellfish before being applied under field conditions. Our results show that shellfish were highly contaminated: enterovirus and HAV RNAs were found in 63 and 67%, respectively, of samples examined with the riboprobes. On the same site, viral (HAV and enterovirus) RNAs were found in a larger fraction of cockles than mussels. Statistical tests of dependence showed no relationship between viral contamination and bacterial contamination (evaluated by fecal coliform counts).
机译:基因组探针用于研究天然床(法国大西洋沿岸)的两种贝类中的甲型肝炎病毒(HAV)和肠病毒RNA。洗脱浓缩后,通过斑点印迹杂交测定通过蛋白酶K提取的核酸并通过苯酚-氯仿纯化的乙醇沉淀。所使用的探针是对应于3'末端(3D聚合酶编码区)的特异性HAV探针和对应于5'非编码区的肠病毒探针。该方法首先在实验条件下使用掺有病毒的贝类进行测试,然后再在野外条件下应用。我们的结果表明,贝类受到高度污染:在用核糖核酸检查的样品中,分别发现肠病毒和HAV RNA的比例分别为63%和67%。在同一位置,发现蛤ussel中的病毒(HAV和肠病毒)RNA比贻贝更大。依赖性的统计检验表明,病毒污染与细菌污染之间没有关系(通过粪便大肠菌群计数评估)。

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