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建立一种重组酶聚合酶扩增检测方法,用于快速检测鼻咽拭子样本中的猪链球菌 2 型。

Development of a recombinase polymerase amplification assay for rapid detection of Streptococcus suis type 2 in nasopharyngeal swab samples.

机构信息

Medical School of Yichun University, Yichun, Jiangxi, China; Jiangxi Provincial Key Laboratory of Active Component of Natural Drugs, Poster-Doctoral Research Center, Jiangxi, China.

Medical School of Yichun University, Yichun, Jiangxi, China.

出版信息

Diagn Microbiol Infect Dis. 2022 Feb;102(2):115594. doi: 10.1016/j.diagmicrobio.2021.115594. Epub 2021 Nov 7.

Abstract

Streptococcus suis serotype 2 (SS2), an emerging zoonotic pathogen, may induce severe infections and symptoms manifested as septicemia, meningitis and even death both in human and pigs. The aim of this article was to develop a new methodology as real-time recombinase polymerase amplification (RT-RPA) assay targeting cps2J gene for the detection of SS2 (or SS1/2). The sensitivity and reproducibility of RT-RPA results were evaluated and compared with a real-time quantitative PCR (RT-qPCR). The established RT-RPA reaction could be completed in 20 minutes with distinguishable specificity against the predominant S. suis infection serotypes of 3, 4, 5, 7, 9, 14, and 31. Lower detection limit for RT-RPA was 10 genomic DNA copies per reaction. The specimen performance of RT-RPA was tested in nasopharyngeal swab samples with the sensitivity and specificity as 97.5% and 100%, respectively. Thus, this RT-RPA method is a rapid and potential molecular diagnostic tool for SS2 detection.

摘要

猪链球菌 2 型(SS2)是一种新兴的人畜共患病病原体,可引起人类和猪严重感染和症状,表现为败血症、脑膜炎,甚至死亡。本文旨在开发一种新的方法,即针对 cps2J 基因的实时重组酶聚合扩增(RT-RPA)检测方法,用于检测 SS2(或 SS1/2)。评估了 RT-RPA 结果的灵敏度和可重复性,并与实时定量 PCR(RT-qPCR)进行了比较。建立的 RT-RPA 反应可在 20 分钟内完成,对主要的猪链球菌感染血清型 3、4、5、7、9、14 和 31 具有可区分的特异性。RT-RPA 的最低检测限为每个反应 10 个基因组 DNA 拷贝。RT-RPA 的标本性能在鼻咽拭子样本中进行了测试,其灵敏度和特异性分别为 97.5%和 100%。因此,该 RT-RPA 方法是一种快速、有潜力的 SS2 检测分子诊断工具。

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