【摘要】：目的建立基于AllGlo探針Real-time RT-PCR技術(shù)檢測發(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片段的相對保守序列設(shè)計引物和AllGlo探針,建立和優(yōu)化Real-time RT-PCR反應(yīng)體系,對其敏感性、特異性和穩(wěn)定性進行評價,并檢測臨床疑似SFTSV病例標本,與普通RT-PCR法、病毒分離法和抗體檢測法進行比較。結(jié)果所建立AllGlo探針Real-time RT-PCR法檢測SFTSV核酸,標準曲線方程為y=-3.38x+46.03(y為Ct值,x為核酸拷貝數(shù)log值),r20.998,擴增效率為97.6%,Ct值變異系數(shù)(CV)均1.2%,檢測限為1.00×103 copy/mL。與漢坦病毒、H1N1、H3N2、登革病毒1~4型均無交叉。檢測362份疑似病例標本,SFTSV核酸陽性檢出率11.9%,高于普通RT-PCR法、病毒分離法和抗體檢測法(P值均0.05)。結(jié)論建立的AllGlo探針Real-time RT-PCR法,操作簡單、快速,靈敏性、特異性較高,可用于SFTSV核酸檢測。
[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：2517994