發(fā)布時(shí)間：2019-07-23 07:19

【摘要】：目的建立基于AllGlo探針Real-time RT-PCR技術(shù)檢測(cè)發(fā)熱伴血小板減少綜合征病毒(SFTSV)的方法。方法根據(jù)SFTSV基因組S片段的相對(duì)保守序列設(shè)計(jì)引物和AllGlo探針,建立和優(yōu)化Real-time RT-PCR反應(yīng)體系,對(duì)其敏感性、特異性和穩(wěn)定性進(jìn)行評(píng)價(jià),并檢測(cè)臨床疑似SFTSV病例標(biāo)本,與普通RT-PCR法、病毒分離法和抗體檢測(cè)法進(jìn)行比較。結(jié)果所建立AllGlo探針Real-time RT-PCR法檢測(cè)SFTSV核酸,標(biāo)準(zhǔn)曲線方程為y=-3.38x+46.03(y為Ct值,x為核酸拷貝數(shù)log值),r20.998,擴(kuò)增效率為97.6%,Ct值變異系數(shù)(CV)均1.2%,檢測(cè)限為1.00×103 copy/mL。與漢坦病毒、H1N1、H3N2、登革病毒1~4型均無(wú)交叉。檢測(cè)362份疑似病例標(biāo)本,SFTSV核酸陽(yáng)性檢出率11.9%,高于普通RT-PCR法、病毒分離法和抗體檢測(cè)法(P值均0.05)。結(jié)論建立的AllGlo探針Real-time RT-PCR法,操作簡(jiǎn)單、快速,靈敏性、特異性較高,可用于SFTSV核酸檢測(cè)。
[Abstract]:Objective to establish a method based on AllGlo probe Real-time RT-PCR for detection of fever complicated with thrombopenia syndrome virus (SFTSV). Methods primers and AllGlo probes were designed according to the relatively conserved sequence of SFTSV genomic S fragment. The Real-time RT-PCR reaction system was established and optimized. The sensitivity, specificity and stability of Real-time RT-PCR reaction system were evaluated. The clinical suspected SFTSV samples were detected and compared with ordinary RT-PCR method, virus isolation method and antibody detection method. Results the standard curve equation for the detection of SFTSV nucleic acid by AllGlo probe Real-time RT-PCR method was y 鈮，

本文編號(hào)：2517994