AllGlo探针Real-time RT-PCR检测发热伴血小板减少综合征病毒

发布时间：2019-07-23 07:19

【摘要】：目的建立基于AllGlo探针Real-time RT-PCR技术检测发热伴血小板减少综合征病毒(SFTSV)的方法。方法根据SFTSV基因组S片段的相对保守序列设计引物和AllGlo探针,建立和优化Real-time RT-PCR反应体系,对其敏感性、特异性和稳定性进行评价,并检测临床疑似SFTSV病例标本,与普通RT-PCR法、病毒分离法和抗体检测法进行比较。结果所建立AllGlo探针Real-time RT-PCR法检测SFTSV核酸,标准曲线方程为y=-3.38x+46.03(y为Ct值,x为核酸拷贝数log值),r20.998,扩增效率为97.6%,Ct值变异系数(CV)均1.2%,检测限为1.00×103 copy/mL。与汉坦病毒、H1N1、H3N2、登革病毒1~4型均无交叉。检测362份疑似病例标本,SFTSV核酸阳性检出率11.9%,高于普通RT-PCR法、病毒分离法和抗体检测法(P值均0.05)。结论建立的AllGlo探针Real-time RT-PCR法,操作简单、快速,灵敏性、特异性较高,可用于SFTSV核酸检测。
[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，

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