B型流感亚系单碱基延伸终止荧光偏振快速检测的研究

Detection of influenza B viruses using single-base extension termination and fluorescence polarization methods

目的建立一种利用单碱基延伸终止结合荧光偏振技术(Fluorescence polarization,FP)快速检测B型流感By(B/Yamagata)和Bv(B/Victoria)亚系的新方法。方法从GenBank下载By和Bv血凝素(HA)基因各50条序列,通过MEGA软件进行分析,利用Primer Premier 5.0设计B型流感病毒亚系特异性引物和单标记延伸引物,建立单碱基延伸终止法与荧光偏振检测方法。结果 40株Bv和40株By病毒用单碱基延伸终止法与荧光偏振检测,与传统的基因测序结果一致性达100%。结论本方法操作简便、敏感性及特异性高、判读结果直观明确、检测费用低,适合于流感监测实验室对流感病毒的快速分子诊断。

Objective To detect influenza B viruses subline by using a novel method based on single-base extension termination and fluorescence polarization. Methods Fifty HA gene sequences coding for B/Yamagata（By） and B/Victoria （By） lineage,respectively,were randomly downloaded form GenBank and analyzed by software MEGA. Primers specific for HA gene of B/Yamagata and B/Victoria lineages were designed by Primer and then applied in the single -base extension termination and fluorescence polarization method developed. Results The results of the detection for the 40 Bv lineage and 40 By lineage by this assay were agreement with that determined by HAI assay. Conclusions The novel method is simpler,high sensitive and specific,inexpensive and the test results were clear, which could be applied in influenza surveiflanee laboratories for rapid molecular diagnosis.