应用RTPCR和RFLP分析肾型鸡传染性支气管炎病毒基因型  

Genotype of Zhejiang isolates of avian infectious bronchitis virus using reverse transcript polymerase chain reaction and restriction fragment length polymorphism analysis:

作　　者：何永强[1] 周继勇[2] 于涟[2] 杜青云[1] 

机构地区：[1]浙江省农业科学院生物技术所,浙江杭州310021 [2]浙江大学动物科学学院,浙江杭州310029

出　　处：《浙江农业学报》2000年第3期117-120,共4页Acta Agriculturae Zhejiangensis

摘　　要：对来自浙江省 3个不同地区的肾型鸡传染性支气管炎病毒 (IBV JD ,DQ ,HY)和参考株H12 0 ,通过蛋白酶K SDS处理、酚 氯仿法抽提基因组RNA ,经RT PCR扩增得到了预期为 1.72kb的S1蛋白基因cDNA ,用限制性内切酶HaeIII,PstI,Ksp632I对S1基因cDNA进行RFLP分析。结果表明JD ,DQ ,HY毒株的RFLP带型相同 ,但与经典的肾型IBV代表株 (即T株、Gray株、Holte株 )以及呼吸型H12 0 均不相同 ,与国内已报道的肾型DL毒株也不同 ,表明浙江省流行的肾型IBV很可能是新的变异株。The genome RNA of three nephropathogenic isolates JD,DQ,HY of avian infectious bronchitis virus isolated from Zhejiang region and one standard strain H 120 were extracted by means of proteinase K/sodium dedecyl sulfate and phenol/chloroform. A 1.72 kb fragment of gene encoding S1 glycopeptide was amplified by reverse transcript polymerase chain reaction and digested with restriction enzyme HaeIII,PstI,Ksp632I.The patterns of restriction fragment length polymorphism analysis showed that isolate JD was the same genotype as isolates DQ,HY, but they differ from other nephropathogenic isolates of IBV reported which included classic nephropathogenic strains T,Gray,Holte, and respiratory strain H 120 .The result indicated that epidemic nephropathogenic isolates of IBV in Zhejiang region were putative variant strains of IBV.

关 键 词：RT-PCR RFLP 肾型鸡传染性支气管炎病毒 基因型 

分 类 号：S858.315.3[农业科学—临床兽医学] S852.65[农业科学—兽医学]