Atmar R L, Neill F H, Woodley C M, Manger R, Fout G S, Burkhardt W, Leja L, McGovern E R, Le Guyader F, Metcalf T G, Estes M K
Department of Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.
Appl Environ Microbiol. 1996 Jan;62(1):254-8. doi: 10.1128/aem.62.1.254-258.1996.
A multicenter, collaborative trial was performed to evaluate the reliability and reproducibility of a previously described method for the detection of Norwalk virus in shellfish tissues with the PCR (R.L. Atmar, F. H. Neill, J. L. Romalde, F. Le Guyader, C. M. Woodley, T. G. Metcalf, and M. K. Estes, Appl. Environ. Microbiol. 61:3014-3018, 1995). Virus was added to the stomachs and hepatopancreatic tissues of oysters or hard-shell clams in the control laboratory, the samples were shipped to the participating laboratories, and viral nucleic acids were extracted and then detected by reverse transcription-PCR. The sensitivity and specificity of the assay were 85 and 91%, respectively, when results were determined by visual inspection of ethidium bromide-stained agarose gels; the test sensitivity and specificity improved to 87 and 100%, respectively, after confirmation by hybridization with a digoxigenin-labeled, virus-specific probe. We have demonstrated that this method can be implemented successfully by several laboratories to detect Norwalk virus in shellfish tissues.
开展了一项多中心协作试验，以评估先前描述的利用聚合酶链反应（PCR）检测贝类组织中诺如病毒方法的可靠性和可重复性（R.L. 阿特马尔、F.H. 尼尔、J.L. 罗马尔德、F. 勒吉亚德、C.M. 伍德利、T.G. 梅特卡夫和M.K. 埃斯蒂斯，《应用与环境微生物学》61:3014 - 3018，1995年）。在对照实验室将病毒添加到牡蛎或硬壳蛤的胃部和肝胰腺组织中，将样本运送到参与试验的实验室，提取病毒核酸，然后通过逆转录PCR进行检测。当通过目视检查溴化乙锭染色的琼脂糖凝胶来确定结果时，该检测方法的灵敏度和特异性分别为85%和91%；在用地高辛配体标记的病毒特异性探针杂交确认后，检测灵敏度和特异性分别提高到87%和100%。我们已经证明，该方法可由多个实验室成功实施，用于检测贝类组织中的诺如病毒。
