开发一种新型数字 RT-PCR 方法，用于检测不同基质中的人杯状病毒。
Development of a novel digital RT-PCR method for detection of human sapovirus in different matrices.
机构信息
Departamento de Microbiología y Parasitología, CIBUS-Facultad de Biología, Universidade de Santiago de Compostela, 15782, Santiago de Compostela, Spain.
Laboratório de Análises, Instituto Superior Técnico, Universidade Técnica de Lisboa, 1049-001, Lisboa, Portugal.
出版信息
J Virol Methods. 2018 Apr;254:21-24. doi: 10.1016/j.jviromet.2018.01.005.
A new nanofluidic digital RT-PCR method was developed for sapovirus (SaV) using control material obtained according to standards for enteric viruses. Primers employed amplify a fragment of 112 bp of the polymerase capsid junction, allowing the detection of human genogroups I, II and IV. Analytical validation was performed in clinical, shellfish and environmental water samples. This novel protocol rendered great effectiveness and repetitiveness, as well as higher sensitivity than real time RT-PCR assay, with differences in quantification ranging from 0.1 to 2.6 log-units. The method described here can constitute a promising tool for standardizing SaV quantification.
一种新的纳米流控数字 RT-PCR 方法被开发出来，用于检测肠道病毒标准物质中获得的肠道病毒（SaV）。所使用的引物扩增聚合酶衣壳连接部的 112bp 片段，可检测到人类基因组 I、II 和 IV。对临床、贝类和环境水样进行了分析验证。与实时 RT-PCR 检测方法相比，该新方案具有更好的有效性和重复性，以及更高的灵敏度，定量差异范围为 0.1 至 2.6 个对数单位。本文描述的方法可以为 SaV 定量标准化提供一种有前途的工具。
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