基于等温 RNA 扩增的荧光检测法对柯萨奇病毒 A16 的新型精确检测方法。

A new accurate assay for Coxsackievirus A 16 by fluorescence detection of isothermal RNA amplification.

机构信息

Laboratory Medicine Center, Children's Hospital of Fudan University, Shanghai 201102, China.

出版信息

J Virol Methods. 2013 Nov;193(2):459-62. doi: 10.1016/j.jviromet.2013.07.013. Epub 2013 Jul 18.

DOI:10.1016/j.jviromet.2013.07.013

Abstract

Coxsackievirus A 16 (CA16) is one of the most common causes of hand, foot, and mouth disease (HFMD) worldwide. Without a vaccine or antiviral drug early, rapid, and accurate detection is critical for preventing and controlling HFMD. A simultaneous amplification and testing (SAT) assay was developed for detecting CA16 based on isothermal RNA amplification with fluorescence using standard, real-time PCR equipment. Primers and probes were designed to target the VP1 region of CA16. Virus strains and clinical specimens were used to evaluate the diagnostic performance characteristics of the assay. The assay detected as few as 10 copies of CA16 RNA transcripts. Using real-time PCR plus sequencing as the reference standard, the sensitivity and specificity of the SAT-CA16 assay were 100% and 99.2%, respectively. These findings indicate that SAT-CA16 is a rapid and reliable method for detecting CA16.

摘要

柯萨奇病毒 A16（CA16）是全球引起手足口病（HFMD）最常见的病原体之一。在没有疫苗或抗病毒药物的情况下，早期、快速和准确的检测对于预防和控制 HFMD 至关重要。本研究基于等温 RNA 扩增与荧光检测，开发了一种用于检测 CA16 的同时扩增和检测（SAT）检测方法，该方法使用标准的实时 PCR 设备。设计了引物和探针以针对 CA16 的 VP1 区域。使用病毒株和临床标本评估了该检测方法的诊断性能特征。该检测方法可检测到低至 10 拷贝的 CA16 RNA 转录本。使用实时 PCR 加测序作为参考标准，SAT-CA16 检测方法的灵敏度和特异性分别为 100%和 99.2%。这些结果表明，SAT-CA16 是一种快速可靠的 CA16 检测方法。