塞内卡谷病毒实时荧光RT-PCR方法的建立

Development of a Real-time RT-PCR Assay for the Detection of Seneca Valley Virus

本研究根据塞内卡谷病毒(SVV)基因保守序列设计特异性引物和探针,建立了检测SVV的实时荧光RT-PCR法。结果表明,该法特异性强,能准确检测SVV核酸,与其他几种相似疾病病原核酸无交叉反应;灵敏度高,本试验中最低可检测核酸浓度为7.2×10^-5μg/mL;重复性好,Ct值批内和批间变异系数(CV)均小于3%。利用所建立方法对116份临床样品进行检测,结果与普通RT-PCR一致。上述结果表明,本研究建立的方法可用于SVV的临床检测和流行病学调查,为有效防控塞内卡病毒病提供技术支撑。

To establish a rapid and specific assay for detection of seneca valley virus(SVV),a real-time fluorescent RT-PCR assay was developed with specific primers and probes designed according to the conservative gene of SVV.The results showed that the assay had high specificity and sensitivity.SVV was detected specifically by the assay without cross-reactions with other inactivated virus antigens.The limit of detection for SVV RNA was 7.2×10^-5μg/mL by the methed.The intra-and inter-assay coefficient of variation of Ct values were less than 3%.A total of 116 clinical samples were tested by the developed assay and general RT-PCR,and the test results were consistent.The real-time fluorescent RT-PCR assay could be used for clinical detection and epidemiological investigation of senecaviral disease.