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基于广泛反应性核酸序列扩增检测法对粪便样本中诺如病毒的检测评估

Evaluation of a broadly reactive nucleic acid sequence based amplification assay for the detection of noroviruses in faecal material.

作者信息

Moore C, Clark E M, Gallimore C I, Corden S A, Gray J J, Westmoreland D

机构信息

National Public Health Service for Wales Microbiology Cardiff, Specialist Virology Centre, University Hospital of Wales, Heath Park, Cardiff CF14 4XW, UK.

出版信息

J Clin Virol. 2004 Apr;29(4):290-6. doi: 10.1016/S1386-6532(03)00170-7.

Abstract

A recently described nucleic acid sequence based amplification (NASBA) assay for the detection of genogroup I (GI) and genogroup II (GII) norovirus RNA in faecal samples was evaluated against a reverse transcription polymerase chain reaction (RT-PCR). Both assays were used to screen a panel of 38 faecal samples known to contain 17 different norovirus strains and 131 clinical samples collected from 60 gastroenteritis outbreaks of unknown aetiology. The NASBA assay detected 13 out of the 17 strains of norovirus in the characterised panel, failing to detect a single GII strain and three GI strains. There was 90% agreement between the two assays used to detect norovirus in clinical samples from outbreaks. NASBA detected norovirus RNA in all 64 samples positive by RT-PCR and also detected norovirus RNA in additional 13 samples that were negative by RT-PCR. The sensitivity and specificity of NASBA was 100% and 80%, respectively, compared to RT-PCR results. The norovirus NASBA assay was shown to be highly sensitive and specific, and its ease of use and rapid turnaround time makes it a favourable alternative to RT-PCR for the investigation of norovirus outbreaks.

摘要

针对一种最近描述的用于检测粪便样本中I基因组(GI)和II基因组(GII)诺如病毒RNA的基于核酸序列扩增(NASBA)的检测方法,与逆转录聚合酶链反应(RT-PCR)进行了对比评估。两种检测方法都用于筛查一组已知含有17种不同诺如病毒株的38份粪便样本以及从60起病因不明的肠胃炎暴发中收集的131份临床样本。NASBA检测方法在特征明确的样本组中检测出了17种诺如病毒株中的13种,未检测出任何一种GII株和3种GI株。在用于检测暴发临床样本中诺如病毒的两种检测方法之间,一致性为90%。NASBA在RT-PCR检测呈阳性的所有64份样本中均检测出诺如病毒RNA,并且在RT-PCR检测呈阴性的另外13份样本中也检测出了诺如病毒RNA。与RT-PCR结果相比,NASBA的灵敏度和特异性分别为100%和80%。诺如病毒NASBA检测方法显示出高度的灵敏性和特异性,并且其易用性和快速周转时间使其成为调查诺如病毒暴发时替代RT-PCR的有利选择。

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