Department of Acute Infectious Diseases Control and Prevention, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing, China.
J Med Virol. 2013 Feb;85(2):370-7. doi: 10.1002/jmv.23415. Epub 2012 Dec 4.
A highly sensitive one-step real-time RT-PCR method using a minor-groove-binding (MGB) probe was developed for detection and quantitation of severe febrile with thrombocytopenia syndrome virus (SFTSV). The assay could discriminate SFTSV infection from other related viral diseases in human with a minimum detection limit of 10 viral RNA copies/µl and was 1,000 times more sensitive than the conventional PCR. Strong linear correlations (r(2) > 0.99) between the C(t) values and viral RNA standards over a linear range were obtained. The coefficients of variation of intra- and inter-assay reproducibility were both less than 2%. The RT-PCR was also shown to be highly specific, as no positive signals were detected for other related viruses. Evaluation of this assay with serum samples from laboratory confirmed cases and healthy donors showed 100% clinical diagnostic sensitivity and over 99% specificity. Clinical application with samples from 287 patients admitted to the hospital with suspected SFTSV infection showed that 15% were infected by SFTSV. This assay was rapid, requiring just over 2 hr, including the nucleic acid extraction step.
建立了一种基于小沟结合（MGB）探针的高灵敏度一步法实时 RT-PCR 方法，用于检测和定量严重发热伴血小板减少综合征病毒（SFTSV）。该检测方法能够区分 SFTSV 感染与人类其他相关病毒疾病，最低检测限为 10 个病毒 RNA 拷贝/µl，比常规 PCR 灵敏 1000 倍。在线性范围内，C(t)值与病毒 RNA 标准品之间具有很强的线性相关性（r(2) > 0.99）。内和间分析重复性的变异系数均小于 2%。该 RT-PCR 也表现出高度的特异性，因为其他相关病毒均未检测到阳性信号。用实验室确诊病例和健康供体的血清样本对该检测方法进行评估，结果显示临床诊断敏感性为 100%，特异性超过 99%。对 287 例因疑似 SFTSV 感染住院的患者样本进行临床应用，结果显示有 15%的患者感染了 SFTSV。该检测方法快速，包括核酸提取步骤在内，只需 2 小时以上。
