摘要
目的建立高致病性禽流感H7和H9亚型荧光定量PCR方法。方法根据禽流感病毒(avian influenzavirus,AIV)HA-H7基因和HA-H9基因分别设计特异AIV H7和H9亚型特异荧光定量PCR引物和探针,优化反应体系和反应条件,分析该试剂的敏感性、特异性、重复性等。结果经优化反应体系和反应条件后,AIV H7试剂和AIV H9试剂的敏感性达到1×104.67EID50和1×103.80EID50水平,特异性与细胞培养敏感性类似;批内和批间精密度Ct值的CV值均远小于10%,可重复性良好。结论本项目研制的AIV H7和H9亚型的敏感性、特异性、重复性均好,适合在禽流感H7和H9亚型疫情和人类疑似禽流感病例中检测使用。
Objective To establish the fluorescence influenza virus （AIV） subtypes H7 and H9. Methods Primers designed for AIV subtypes H7 and H9, respectively, according quantitative PCR method for detection of avian and probe of fluorescence quantitative PCR were to the AIV HA H7 and H9 genes. The reaction system and conditions were optimized. Sensitivity, specificity and reproducibility of the system were analyzed. Results After optimization of the reaction system and its conditions, the sensitivity of H7 and H9 assay reached 1 10^4.67 EID50 and 1 10^3.80 EID50 respectively, and their specificity was similar to that of cell culture. The CV values of CT in intra-and inter-batch were far less than 10%, showing that their reproducibility was excellent. Conclusion The sensitivity, specificity and reproducibility of AIV subtypes H7 and H9 are rather good. AIV subtypes H7 and H9 can be used in detection of avian epidemic in suspected avian H7 and H9- infected human cases.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2010年第3期246-249,共4页
Journal of Third Military Medical University
基金
国家自然科学基金(30972757)
广东省科技项目(2004A2090102)~~
