H1、H3、H9亚型禽流感病毒多联RT-PCR检测方法的建立

Development of RT-PCR detection method for differentiation of H1,H3 and H9 subtype avian influenza virus simultaneously

根据GenBank中登录的H1、H3、H9亚型禽流感病毒(AIV)血凝素(HA)基因序列,通过多序列对比,在保守区域内设计了3对特异性引物,对PCR反应体系的各组分进行试验调整,建立了一种H1、H3、H9亚型AIV多联RT-PCR检测方法,在2h内即可判断待检样品中是否有这3种亚型AIV的存在。经灵敏性及特异性试验以及临床样品检测结果证明,该方法灵敏度高、特异性好、结果准确,能够作为H1、H3、H9亚型AIV的快速检测方法。

Based on the sequence of hemagglutinin （HA） gene of H1, H3 and H9 subtype avain in- fluenza virus （AIV） in GenBank,we designed three pairs of specific differential primers in the con- served region via multiple sequence comparison,then a RT-PCR system for differentiation of HI, H3 and H9 subtype AIV was development. The RT-PCR system can simultaneously differentiate H1 ,H3 and H9 AIV of clinical samples within 2 hours ; furthermore, the system has highly sensi- tivity,specificity and accuracy compared to the conventional RT-PCR methods. In conclusion, the developed RT-PCR system is a rapid detection method for differentiation of H1, H3 and H9 AIV of clinical samples at the same time.