豬傳染性胃腸炎病毒實(shí)時(shí)熒光定量PCR檢測(cè)方法的建立及應(yīng)用
發(fā)布時(shí)間：2018-11-20 08:13
【摘要】：針對(duì)豬傳染性胃腸炎病毒(TGEV)S基因的保守序列設(shè)計(jì)引物,以TEGV疫苗毒株為模板,克隆S基因,并構(gòu)建重組陽(yáng)性質(zhì)粒,優(yōu)化反應(yīng)體系和擴(kuò)增條件,建立基于SYBR GreenⅠ染料的實(shí)時(shí)熒光定量PCR檢測(cè)方法,并對(duì)其特異性、重復(fù)性和靈敏度進(jìn)行分析。結(jié)果顯示:建立的檢測(cè)方法靈敏度可達(dá)10拷貝/μL,標(biāo)準(zhǔn)曲線線性關(guān)系良好(r=0.997),擴(kuò)增效率高,具有良好的特異性和重復(fù)性。使用該方法對(duì)124份臨床疑似TGEV病料進(jìn)行檢測(cè),陽(yáng)性樣本有17份,檢測(cè)樣本的陽(yáng)性率為13.7%。該方法可用于獸醫(yī)臨床上TGEV的快速檢測(cè)和流行病學(xué)分析。
[Abstract]:According to the conserved sequence of (TGEV) S gene of porcine transmissible gastroenteritis virus, the S gene was cloned by using TEGV vaccine strain as template, and the recombinant positive plasmid was constructed to optimize the reaction system and amplification conditions. A real-time fluorescent quantitative PCR method based on SYBR Green 鈪，
本文編號(hào)：2344360
[Abstract]:According to the conserved sequence of (TGEV) S gene of porcine transmissible gastroenteritis virus, the S gene was cloned by using TEGV vaccine strain as template, and the recombinant positive plasmid was constructed to optimize the reaction system and amplification conditions. A real-time fluorescent quantitative PCR method based on SYBR Green 鈪，
本文編號(hào)：2344360
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