鸡传染支气管炎病毒H52、H120和M41株的PCR鉴别方法研究

Identification of Avian Infectious Bronchitis Virus Strains H120,H52 and M41 with PCR Method

根据GenBank发表的序列,对鸡传染性支气管炎病毒(IBV)H52、H120和M41株全基因组序列进行比较,设计并合成了两对特异性引物,其中一对引物能以M41株为模板,特异性扩增出1791 bp的目的片断,从而特异鉴定IBV M41株;另一对引物能以H52株和H120株为模板,特异性扩增1700 bp的目的片断,再用限制性内切酶NaeⅠ对PCR产物进行酶切,H120株能够被切成1000 bp和700 bp两个片段,而H52株的PCR产物不存在该酶切位点,从而能区分H52株和H120株。本研究建立的PCR方法和技术具有快速、简单、特异性强等优点,可用于IBV H52、H120和M41株活疫苗的分子鉴别。

Based on GenBank published sequences and analysis to whole genome sequences of three avian infectious bronchitis virus（IBV） strains H52, H120 and M41, two pairs of specific primers were designed and synthesized. Using one pair primers, the target fragment of 1791 bp of IBV M41 strain was amplified to be identified specially. The other pair primers could amplify the target fragment of 1700 bp of strains H52 and H120. Then the PCR products were digested by restriction endonuclease Nae I , the PCR products of H120 can be cut into two fragments of 1000 bp and 700 bp, but the PCR products of strain 1-152 could not be cut since there is no Nae I restriction site. Thus this method was used to distinguish between strain H52 and strain H120. The method was fast, simple and specific, and could be used for the molecular identification of IBV strains H52, H120 and M41.