通过逆转录半巢式聚合酶链反应检测天然污染贝类和沉积物中的甲型肝炎病毒、轮状病毒和肠道病毒。
Detection of hepatitis A virus, rotavirus, and enterovirus in naturally contaminated shellfish and sediment by reverse transcription-seminested PCR.
作者信息
Le Guyader F, Dubois E, Menard D, Pommepuy M
机构信息
Laboratoire de Microbiologie, Institut Français de Recherche pour l'Exploitation de la Mer, Nantes, France.
出版信息
Appl Environ Microbiol. 1994 Oct;60(10):3665-71. doi: 10.1128/aem.60.10.3665-3671.1994.
A reverse transcription-PCR method was developed to detect enterovirus (EV), hepatitis A virus (HAV), and rotavirus (RV) RNAs in shellfish and sediment. The method was first tested under experimental conditions by using virus-spiked shellfish to evaluate assay sensitivity. The use of CC41 cellulose was found to be efficient for removing inhibitors of RV detection. For sediment samples, a Sephadex column was used to allow the detection of EV and HAV RNAs. The specificity of amplified products was controlled by hybridization with digoxigenin-labeled oligoprobes. The method was then applied to naturally contaminated shellfish and sediments. EV, HAV, and RV RNAs were detected in 22, 14, and 20% of the shellfish samples, respectively. No relationship between viral contamination and bacterial contamination was found. When viral RNAs (HAV or EV) were detected in sediments, they were also detected in shellfish.
开发了一种逆转录聚合酶链反应（RT-PCR）方法，用于检测贝类和沉积物中的肠道病毒（EV）、甲型肝炎病毒（HAV）和轮状病毒（RV）RNA。该方法首先在实验条件下，通过使用接种病毒的贝类来评估检测灵敏度。发现使用CC41纤维素可有效去除RV检测的抑制剂。对于沉积物样本，使用葡聚糖凝胶柱以检测EV和HAV RNA。通过与地高辛标记的寡核苷酸探针杂交来控制扩增产物的特异性。然后将该方法应用于天然污染的贝类和沉积物。在分别22%、14%和20%的贝类样本中检测到了EV、HAV和RV RNA。未发现病毒污染与细菌污染之间存在关联。当在沉积物中检测到病毒RNA（HAV或EV）时，在贝类中也检测到了这些病毒RNA。
Zotero 插件