基于SH片段的中国腮腺炎病毒疫苗株与野毒株鉴别方法的建立

The establishment of method for identifying China vaccine strains and wild strains of Mumps virus based on the SH fragment

目的研究建立一种简便快速的鉴别中国腮腺炎病毒疫苗株与野毒株方法。方法在已扩增出的小疏水蛋白基因(small hydrophobic gene)(SH基因)上,寻找能将中国腮腺炎病毒疫苗株区别于野毒株的限制性内切酶酶切位点,并对逆转录-聚合酶链反应(Reverse Transcription-Polymerase Chain Reaction,RT-PCR)的方法进行特异性实验证明。结果非腮腺病毒RT-PCR结果均未见阳性条带,说明RT-PCR方法特异。PCR产物经BclI酶切作用后,腮腺炎野毒株将会被酶切为170bp和340bp两个片段,而疫苗株则被酶切为270bp和240bp两个片段。结论新建立的RT-PCR-RFLP是一种快速、简便的鉴别中国腮腺炎病毒疫苗株与野毒株的方法。

Objective To establish a simple and quick method for identifying China vaccine strains and wild strains of Mumps Virus. Methods To search the enzyme site in Small Hydrophobic gene of mumps virus for different domestic vaccine strains and wild strains of mumps virus and then to confirm the specificity of the RT-PCR method,and then to identify the RT-PCR product by RFLP. Results No positive bands can be found in the non-mumps virus strains,it means that the RT-PCR method has good specificity,the PCR products of China mumps vaccine strains of S79 and Jeryl-lynn were all cut into two fragments（270bp and 240bp） by Bcl I,but mumps wild virus strains were all cut into two fragments（170bp and 340bp） by BclI. Conclusion The RTPCR-RFLP method we established is a rapid and simple method for identifying China vaccine strain and wild strain of mumps virus.