利用焦磷酸测序技术检测H7N9禽流感病毒NA基因分子标签

Detection of An NA Gene Molecular Marker in H7N9 Subtype Avian Influenza Viruses by Pyrosequencing

本研究旨在建立H7N9亚型禽流感病毒NA基因分子标签的焦磷酸测序检测方法,用于H7N9型禽流感病毒的快速检测和鉴定。通过对公开发表的H7N9亚型禽流感病毒NA基因序列进行比对,发现分离株在NA基因特定区域缺失的15个核苷酸可作为H7N9亚型禽流感病毒NA基因特异性的分子标签。通过设计覆盖此区域的特异性扩增引物和测序引物,建立了H7N9型禽流感病毒NA基因焦磷酸测序检测方法。基于NA基因特定缺失区域建立的焦磷酸测序技术能用于H7N9亚型禽流感病毒国内分离株NA基因的快速检测和鉴定,且具有较好的特异性和重复性。通过对3株H7N9亚型禽流感病毒分离株的检测,表明3株H7N9亚型禽流感病毒的NA基因均存在15个核苷酸缺失的分子标签。本研究建立的H7N9亚型禽流感病毒NA焦磷酸测序检测方法,可用于H7N9禽流感病毒的检测和鉴定。

This study aimed to establish a method for the detection and identification of H7N9avian influenza viruses based on the NA gene by pyrosequencing.According to the published NA gene sequences of the avian influenza A（H7N9）virus,a 15-nt deletion was found in the NA gene of H7N9avian influenza viruses.The 15-nt deletion of the NA gene was targeted as the molecular marker for the rapid detection and identification of H7N9avian influenza viruses by pyrosequencing.Three H7N9avian influenza virus isolates underwent pyrosequencing using the same assay,and were proven to have the same 15-nt deletion.Pyrosequencing technology based on the NA gene molecular marker can be used to identify H7N9avian influenza viruses.