基孔肯雅病毒的实时荧光RT-PCR检测方法研究  

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

作　　者：黄吉城[1] 郑夔[1] 李小波[1] 洪烨[1] 师永霞[1] 幸芦琴[1] 相大鹏[1] 郭波旋[1] 钟玉清[1] 

机构地区：[1]广东出入境检验检疫局检验检疫技术中心,广州510623

出　　处：《中国卫生检验杂志》2008年第9期1721-1723,共3页Chinese Journal of Health Laboratory Technology

基　　金：国家质量监督检验检疫总局行业标准制定项目资助(2007B198)

摘　　要：目的:建立基孔肯雅病毒的荧光定量PCR检测方法。方法:人工合成基孔肯雅病毒部分有代表性的序列核酸作为阳性对照模板,设计几组实时荧光PCR引物、探针及反应体系,摸索出最佳反应体系条件。结果:建立的基孔肯雅病毒的实时荧光PCR检测方法最佳反应体系为:正向引物CH IK-FP终浓度500 nM,反向引物CHIK-RP终浓度1000 nM,探针CHIK-Probe终浓度250 nM;扩增程序为:50℃10 m in,95℃10 min,95℃10 s→62℃30 s 45次循环。结论:该方法灵敏度高、特异性强,适用于基孔肯雅病毒的快速检验。Objective：To set up real time RT-PCR detection method for Chikungunya virus.Methods：Synthesizing some representative nucleic acid segment of Chikungunya virus as positive control,designing several primers,probes and reaction system of real time RT-PCR to explore the best detection condition.Results：The best detection condition of real time RT-PCR for Chikungunya virus were as following： forward primer concentration 500 nM,reverse primer concentration 1000 nM,detection probe concentration 250 nM,amplifying sequence： 50℃ 10 min,95℃ 10 min,95℃ 10 s→62℃ 30 s 45cycles.Conclusion：This method is suitable for laboratory detection of Chikungunya virus because of its high sensitivity and specificity.

关 键 词：基孔肯雅病毒 实时荧光PCR 检测 

分 类 号：Q78[生物学—分子生物学]