马尔堡病毒的实时荧光RT-PCR检测方法研究

Study on real time RT-PCR detection method for Marburg virus

目的建立马尔堡病毒的实时荧光RT-PCR检测方法。方法人工合成马尔堡病毒特异性核酸序列作为阳性对照模板,设计实时荧光RT-PCR引物、探针并构建反应体系,对反应条件进行优化,验证该方法的特异性、灵敏度。结果建立的马尔堡病毒实时荧光RT-PCR检测方法对马尔堡病毒核酸检测有高度特异性,与1型～4型登革病毒、日本脑炎病毒和基孔肯雅病毒均无交叉反应,检测灵敏度为102拷贝/反应。结论该方法灵敏度高、特异性强,适用于对马尔堡病毒的快速检验。

Objective To set up real time RT-PCR detection method for Marburg virus.Methods Some representative nucleic acid segment of Marburg virus as positive control were synth esized,and several primers,probes and reaction system of real time RT-PCR were designed to explore the best detection condition.The PCR condition was optimized to improve the sensitivity and specificity of the assay.Results The specificity of the assay for real time RT-PCR for Marburg virus was high and there was no cross reactions with Dengue virus,Japanese encephalitis virus and Chikungunya virus.The sensitivity of the assay was 100 gene copies per test.Conclusion This method is suitable for laboratory detection of Marburg virus because of its high sensitivity and specificity.