刘胜牙1,朱玉兰1,胡孔新2,谢聪贤1,甄胜西1,徐媛1
1.深圳国际旅行卫生保健中心,广东 深圳 518033;2.中国检验检疫科学研究院
摘要:目的 建立一种快速、灵敏、高效的人冠状病毒HKU1(HCoV-HKU1)LAMP扩增检测方法。方法 应用生物信息学软件设计5套LAMP扩增引物,以合成的HCoV-HKU1质粒为模板,通过实时浊度法进行最佳引物组的筛选,并分析该方法的特异性和灵敏度。结果 HCoV-HKU1LAMP扩增方法针对其他病原微生物及人类基因组无反应信号。该方法可以检测到1拷贝/?滋l的HCoV-HKU1质粒。结论 HCoV-HKU1 LAMP扩增方法特异性好、灵敏度高,在临床鉴别诊断和口岸冠状病毒的检测中具有很好的应用前景。
关键词:环介导等温扩增;人冠状病毒HKU1;检测;研究
中图分类号:R446.6 文献标识码:B
Study of loop-mediated isothermal amplification technique fordetection of HCoV-HKU1
LIU Sheng-ya*, ZHU Yu-lan, HU Kong-xin, XIE Cong-xian, ZHENSheng-xi, XU Yuan
*Shenzhen International Travel Healthcare Center, Shenzhen,Guangdong 518033, China
Abstract: Objective To establish a rapid detection methodof human coronavirus HKU1 (HCoV-HKU1) based on loop-mediatedisothermal amplification (LAMP). Methods With the applicationof LAMP, five sets of LAMP primers were designed. The synthesizedHCoV-HKU1 gene was used as a template, and the optimal primers wereset by real-time turbidity method. Specificity and sensitivity ofthe method were assessed. Results Other pathogenic organismsand human genome in the method have not shown any response signal.In the experiment of sensitivity, HCoV-HKU1 plasmid gene hada detection limit of 1 copy/?滋l in each reaction. Conclusion This study indicates that our LAMP method is a rapid,sensitive, specific method to detect pathogen of HCoV-HKU1, and hasa good application prospect in clinical differential diagnosis andcoronavirus monitoring of port.
Key words: LAMP; HCoV-HKU1; Detection; Study
《中国国境卫生检疫杂志》2014年4期