摘要
目的利用荧光定量RT-PCR技术建立一种快速检测A型流感病毒的方法。方法根据A型流感M基因的相对保守序列分别设计一对引物及其相应的Taqman探针,建立、优化反应体系后,利用10倍稀释法检验方法的灵敏度并建立相对定量标准曲线;特异性检验后利用临床标本与传统的血凝抑制法进行比较。结果A型流感病毒检测反应的灵敏度为2.56×10-6TCID50,标准曲线相关系数为0.999,扩增效率为108.5%,5种非A型流感病毒病原体检测均为阴性,说明此方法只有很好的稳定性、重现性和特异性。结论本研究建立荧光定量RT-PCR技术可以准确检测A型流感病毒,不仅灵敏度高、稳定性好,而且可以对病毒滴度进行定量检测。
Objectlve To develop a method for rapid detection of influenza virus A by real - time quantitative RT - PCR, Methods Based on the M gene of influenza virus A, a set of primers and Taqman probe were designed. After the quantitative curve of the assay were established using tenfold serial dilution of TCID50, the sensitivity and the specificity were determined. Results For the influenza virus A, the sensitivity of the assay was 5.0×10^-6 TCID50 and the regression coefficient of the quantitative curve was 0.999. Specificity was determined by testing five other specimens, all of which yielded negative results. Conclusion The detection system based on real- time RT- PCR was rapid, sensitive and stable and the viral load can also be determined.
出处
《中国热带医学》
CAS
2006年第1期33-35,共3页
China Tropical Medicine
基金
深圳市科技局计划项目(JH200507131122A)
关键词
A型流感病毒
荧光定量RT—PCR
快速检测
Influenza virus A
Fluorescent Quantitative RT- PCR
Rapid Detection
Standard Curve
