豬流行性腹瀉病毒環(huán)介導(dǎo)等溫擴(kuò)增檢測方法的建立及應(yīng)用
發(fā)布時間：2018-12-07 10:27
【摘要】：試驗旨在建立一種快速檢測豬流行性腹瀉病毒(PEDV)的環(huán)介導(dǎo)等溫擴(kuò)增方法(LAMP),為診斷PEDV提供簡便、敏感、準(zhǔn)確可靠的工具。參考GenBank中PEDV基因序列(登錄號:KT799997),針對PEDV N基因設(shè)計了6條引物,對所建立的LAMP反應(yīng)體系、反應(yīng)溫度進(jìn)行優(yōu)化,建立可特異性擴(kuò)增PEDV的LAMP方法。結(jié)果顯示,本試驗成功建立了PEDV LAMP檢測方法,在60℃恒溫下反應(yīng)60min,能特異性地檢測PEDV,檢測限量為91拷貝/μL,比常規(guī)PCR方法的敏感性高100倍。對比75份臨床樣本的LAMP和常規(guī)RT-PCR法檢測結(jié)果,顯示兩種方法符合率為97.3%。綜上所述,本試驗建立的LAMP方法具有特異性強(qiáng)、敏感性高,操作簡單,設(shè)備要求低的特點,適用于PEDV臨床樣本的快速檢測。
[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，
本文編號：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，
本文編號：2367059
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