诺如病毒快速检测方法的建立及其在国境卫生检疫中的应用

Development of a fast method for detecting norovirus and its application at port

目的:建立快速、特异的诺如病毒实时荧光RT-PCR与常规RT-PCR相结合检测方法,对人感染诺如病毒进行快速检测。方法:根据国外最新流行的诺如病毒核酸序列,设计并合成诺如病毒基因扩增引物及荧光标记探针,摸索最佳反应条件,制定出快速、灵敏检测诺如病毒核酸的实时荧光RT-PCR检测方法及常规RT-PCR检测方法。结果:实验证明,本方法将实时荧光RT-PCR与常规RT-PCR检测方法相结合,能在5 h内快速确认诺如病毒,检测灵敏度达到102拷贝/反应,具有快速、灵敏的特点,且具有较高的特异性和重复性。结论:本实验室利用本方法,首次在广州某国境口岸从外籍发热船员样本中检出5例输入性GGⅡ群诺如病毒。该方法对防止诺如病毒传入我国,保障我国国境卫生安全具有重要意义。

Objective：To establish a method of real - time fluorescent RT - PCR and routine RT - PCR for detecting Norovirus. Methods ：The specific primers and probes were designed based on large scale norovirus genome consensus analysis, a TaqMan - based real - time RT - PCR and a RT - PCR assay were established for detection of norovirus RNA. Results： The method showed high sensitivity （102copies/reaction）, specificity and repetition character, which can affirm nucleic acid of norovirus accurately within five hours. Conclusion：Norovirus Genogroup Ⅱ positive from 11 samples of a foreign ship at port by this method. It is the first time to detect Norovirus from foreign shipping crew by the Bureau of Inspection and Quarantine in China. This method is suit for the detection of norovirus and it has the vital significance of preventing norovirus spreading to our country and safeguarding the health security of our country.