绵羊肺腺瘤病毒套式RT-PCR检测方法的建立

Establishment of a nest RT-PCR method for detecti on of Jaagsiekte sheep retrovirus

根据GenBank中的绵羊肺腺瘤病毒(JSRV)基因组序列,应用Oligo6.0软件设计合成了2对引物,外部引物的扩增片段大小为467bp,内部引物的扩增片段大小为214bp,建立了一种快速检测JSRV的套式RT-PCR检测方法。试验结果显示,该检测方法与同属的几种绵羊易感的病毒均无交叉反应,比普通RT-PCR方法敏感性高,具有重复性好、特异性强、敏感性高等优点,可以准确快速地检测出极低含量的JSRV病原,为绵羊肺腺瘤病的快速诊断、净化及深入研究奠定了基础。

Based on the genome sequence of Jaagsiekte sheep retrovirus（JSRV）available in G enBank,two pairs of primers were designed using Oligo6.0 biology software,the ou ter primers amplified a fragment of 467 bp in length,and the inner primers ampl ification fragment size was 214 bp in length,and a nest RT-PCR detection metho d for JSRV was established.There were no cross reaction with both homologous and sheep susceptible viruses,and the sensitivity is higher than the conventional RT-PCR methods.This method had good reproducibility,specificity and sensitivity,and might detec t low content JSRV accurately and rapidly.This method could be used for rapid diagnosis of JSRV and helpful for eliminatio n of the disease.