传染性支气管炎病毒山东及北京分离株血清型及RFLP基因分型的比较研究

Comparing the serotype and genotype of local isolates of IBV from Shandong and Beijing by RFLP

分离并鉴定出山东省不同地区的 IBV分离株 A、B、C、D和北京分离株 F,利用中和试验分别对各分离株及标准株M4 1 、T等进行血清分型。设计一对引物 ,分别对各毒株的 S1 基因进行 RT- PCR扩增 ,扩增片段长约 170 0 bp,利用其 PCR产物进行 Hae 酶切 ,根据酶切图谱 ,进行基因分型。结果表明 :A、C、D和 M4 1 酶切图谱相同 ,属 Mass基因型 ,分别为 90 0、4 0 0和30 0 bp左右的片段 ;北京 F株有 4条酶切条带 ,与 4 /91基因型不同 ,分别为 10 0 0、30 0、2 0 0和小于 10 0 bp左右的片段 ;B株有4条酶切条带 ,理论推断与 T株相似 ,分别为 70 0、5 0 0、30 0和 180 bp左右的片段 ,可能为同一基因型。血清学分型结果与基因分型结果不符。

Isolated and Identified IBV 5 local isolates (A,B,C,D from Shandong and F from Beijing), Cross neutralization tests were performed in tracheal organ cultures(TOC)to compare the serotype of IBV M 41 ,T and those isolates. The result of serotyping showed that A and T, B and D, F and 4/91 were the same serotype. A pair of primers of S1 gene to those IBV local isolates were designed according to strain M 41 (Gene Bank NO. M21883). S1 genes of them were amplified by RT PCR. PCR products were digested with restriction endonuclease HaeⅢ, and the RELP results showed that A, C, D, M 41 shared the same RFLP pattern, there were 900?400 and 300bp segments, whereas F or B had different RFLP pattern. A, C, D, M 41 were genotype of Mass; F was alone, there were 1000?300?200 and 100bp segments. There were 700?500?300 and 180bp segments in B, B maybe share with genotype of T in theory; The results of serotype and genotype showed maybe some difference in them.