猪流行性腹泻病毒环介导等温扩增检测方法的建立及应用
发布时间：2018-12-07 10:27
【摘要】：试验旨在建立一种快速检测猪流行性腹泻病毒(PEDV)的环介导等温扩增方法(LAMP),为诊断PEDV提供简便、敏感、准确可靠的工具。参考GenBank中PEDV基因序列(登录号:KT799997),针对PEDV N基因设计了6条引物,对所建立的LAMP反应体系、反应温度进行优化,建立可特异性扩增PEDV的LAMP方法。结果显示,本试验成功建立了PEDV LAMP检测方法,在60℃恒温下反应60min,能特异性地检测PEDV,检测限量为91拷贝/μL,比常规PCR方法的敏感性高100倍。对比75份临床样本的LAMP和常规RT-PCR法检测结果,显示两种方法符合率为97.3%。综上所述,本试验建立的LAMP方法具有特异性强、敏感性高,操作简单,设备要求低的特点,适用于PEDV临床样本的快速检测。
[Abstract]:The purpose of this study was to establish a rapid detection method for porcine epidemic diarrhea virus (PEDV) by (LAMP), which provides a simple, sensitive, accurate and reliable tool for the diagnosis of PEDV. Referring to the sequence of PEDV gene in GenBank (accession number: KT799997), six primers were designed for PEDV N gene. The LAMP reaction system and reaction temperature were optimized, and the LAMP method for specific amplification of PEDV was established. The results showed that the PEDV LAMP assay was successfully established, and the detection limit of PEDV, was 91 copies / 渭 L at 60 鈩，
本文编号：2367059
[Abstract]:The purpose of this study was to establish a rapid detection method for porcine epidemic diarrhea virus (PEDV) by (LAMP), which provides a simple, sensitive, accurate and reliable tool for the diagnosis of PEDV. Referring to the sequence of PEDV gene in GenBank (accession number: KT799997), six primers were designed for PEDV N gene. The LAMP reaction system and reaction temperature were optimized, and the LAMP method for specific amplification of PEDV was established. The results showed that the PEDV LAMP assay was successfully established, and the detection limit of PEDV, was 91 copies / 渭 L at 60 鈩，
本文编号：2367059
本文链接：https://www.wllwen.com/yixuelunwen/dongwuyixue/2367059.html
