实时逆转录重组酶聚合酶扩增法快速检测猪流行性腹泻病毒。
Real-time reverse transcription recombinase polymerase amplification assay for rapid detection of porcine epidemic diarrhea virus.
机构信息
Hebei Academy of Science and Technology for Inspection and Quarantine, Shijiazhuang 050051, China; Center of Inspection and Quarantine, Hebei Entry-Exit Inspection and Quarantine Bureau, Shijiazhuang 050051, China.
Hebei Animal Disease Control Center, Shijiazhuang 050050, China.
出版信息
J Virol Methods. 2018 Mar;253:49-52. doi: 10.1016/j.jviromet.2018.01.001. Epub 2018 Jan 9.
Porcine epidemic diarrhea (PED), which is caused by porcine epidemic diarrhea virus (PEDV), is an acute, highly contagious enteric disease characterized by severe watery diarrhea, vomiting, dehydration, and high mortality in suckling piglets. A real-time reverse-transcription recombinase polymerase amplification assay (RT-RPA) was developed based on the nucleocapsid gene of PEDV. RT-RPA assay was performed at 40 °C for 20 min. The assay could detect both the classical and variant PEDV strains, and there was no cross-reaction with other pathogens tested. Using the in vitro transcribed PEDV RNA as template, the analytical sensitivity was 23 copies per reaction. The assay performance was evaluated by testing 76 clinical samples. PEDV RNA positive rate was 55.3% (42/76) by RT-RPA and 59.2% (45/76) by real-time RT-PCR. The diagnostic agreement between the two assays was 96.1% (73/76), and the R value of the two assays was 0.903 by linear regression analysis. The developed RT-RPA assay provides a useful alternative tool for simple, rapid and reliable detection of PEDV in resource-limited diagnostic laboratories and on-site facilities.
猪流行性腹泻（PED）由猪流行性腹泻病毒（PEDV）引起，是一种以仔猪严重水样腹泻、呕吐、脱水和高死亡率为特征的急性、高度传染性肠道疾病。本研究基于 PEDV 的核衣壳基因，建立了一种实时逆转录重组酶聚合酶扩增检测（RT-RPA）方法。RT-RPA 检测在 40°C 下进行 20 分钟。该检测方法可以同时检测经典和变异 PEDV 株，与检测的其他病原体无交叉反应。使用体外转录的 PEDV RNA 作为模板，分析灵敏度为 23 拷贝/反应。通过检测 76 份临床样本评估了检测方法的性能。PEDV RNA 阳性率为 RT-RPA 检测 55.3%（42/76），实时 RT-PCR 检测 59.2%（45/76）。两种检测方法的诊断一致性为 96.1%（73/76），线性回归分析两种检测方法的 R 值为 0.903。本研究建立的 RT-RPA 检测方法为资源有限的诊断实验室和现场设施中简单、快速、可靠地检测 PEDV 提供了一种有用的替代工具。
Zotero 插件