辛德比斯病毒的实时荧光RT-PCR检测方法研究  

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

作　　者：洪烨[1] 袁帅[1] 戴俊[1] 郑夔[1] 黄吉城[1] 师永霞[1] 李小波[1] 苏锦坤[1] 

机构地区：[1]广东出入境检验检疫局检验检疫技术中心,广东广州510623

出　　处：《中国国境卫生检疫杂志》2015年第3期165-167,共3页Chinese Journal of Frontier Health and Quarantine

基　　金：广东检验检疫局科技计划项目(2013GDK35)

摘　　要：目的建立辛德比斯病毒的实时荧光RT-PCR检测方法。方法人工合成辛德比斯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。结果建立的辛德比斯病毒实时荧光RT-PCR检测方法对辛德比斯病毒核酸检测有高度特异性,与1~4型登革病毒、流行性乙型脑炎病毒和基孔肯雅病毒均无交叉反应,检测灵敏度为103拷贝/反应。结论该方法灵敏度高、特异性强,适用于对辛德比斯病毒的快速检验。Objective To set up a real time RT-PCR method for detecting Sindbis virus. Methods Synthesizing the representative nucleic acid segments of Sindbis virus genome as a positive control, designing primers, probes and reaction system of real time RT-PCR 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 Sindbis virus was high and there were no cross reactions with Dengue virus, Japanese encephalitis virus and Chikungunya virus. The sensitivity of the assay was 1 000 gene copies per test. Conclusion This method is suitable for laboratory detection of Sindbis virus because of its high sensitivity and specificity.

关 键 词：辛德比斯病毒 实时荧光RT-PCR 检测 

分 类 号：R512.8[医药卫生—内科学]