重组酶聚合酶扩增联合侧流试验快速检测致病性猪链球菌2型

Yong Qi, Wei Li, Xiaoling Li, Wanpeng Shen, Rui-Chen Lv, Nianhong Lu, Jiameng Li, Susu Zhuang, Yingjia Xu, Qiyuan Gui, Hongbing Shen, Yuexi Li
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引用次数: 1

摘要

猪链球菌血清2型(SS2)是一种重要的人畜共患病原体,可引起猪和人的严重疾病甚至死亡。SS2造成的公共卫生事件和经济损失引起了广泛关注。由于无法获得疫苗,需要开发快速检测方法,及时诊断SS2感染或受污染产品,并监测其在易感动物和人群中的流行情况,以帮助预防和控制SS2感染。设计并合成了一套针对cpsJ2基因的重组酶聚合酶扩增(RPA)引物和探针。侧流(LF)试验结合RPA提供视觉结果。筛选扩增效率高的引物,并对反应体系进行优化。评估检测有效性指标。所建立的方法对SS2的检测限为100拷贝/反应。在受感染的动物样本中,敏感性为100%。检测可在20分钟内完成,只需要恒温设备。所建立的快速、直观、灵敏、特异的RPA-LF检测方法具有较好的检测效果,有望在资源有限地区广泛应用于抗SS2感染。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Rapid Visual Detection of Pathogenic Streptococcus suis Type 2 through a Recombinase Polymerase Amplification Assay Coupled with Lateral Flow Test
Streptococcus suis serotype 2 (SS2) is an important zoonotic pathogen causing serious disease and even death in pigs and humans. Public health events and economic losses caused by SS2 have prompted widespread concern. Because of the unavailability of vaccines, the development of rapid detection methods for timely diagnosis of SS2 infection or contaminated products, and monitoring of its prevalence in susceptible animals and populations, is required to aid in the prevention and control of SS2 infections. Several sets of primers and one probe for a recombinase polymerase amplification (RPA) assay targeting the cpsJ2 gene were designed and synthesized. Lateral flow (LF) tests in combination with RPA were used to provide visual results. Primers with high amplification efficiency were screened, and the reaction system was optimized. Indicators of detection effectiveness were evaluated. The established method had a detection limit of 100 copies/reaction for recognizing SS2 rather than other organisms. The sensitivity was 100%, as evaluated in infected animal samples. The detection could be completed within 20 min and required only constant temperature equipment. The established rapid, visual, sensitive and specific RPA-LF assay showed superior detection performance and is expected to be widely applied to fight SS2 infection in resource-limited areas.
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