鸡传染性法氏囊病病毒RT-PCR检测方法的建立

根据GenBank中已经发表的传染性法氏囊病病毒（IBDV）VP2基因的高度保守序列，设计并合成了一对引物，Blast在线检索GenBank数据库，未发现其他相似序列。提取已鉴定的IBDV—QD株传代病毒尿囊液的总RNA，合成cDNA模板，优化RT-PCR反应条件，最终获得预期453bp的目的片段，IBDV扩增产物经测序结果证实与GenBank中已发表的致病力不同的IBDV毒株相应序列同源性达97．4％～99．9％，最终建立了RT—PCR检测方法。用已建立的RT—PCR方法对已知的8份临床IBDV阳性法氏囊病料进行检测，阳性率达100％，另外对2份鸡白血病病毒（ALDV），2份鸡传染性喉气管炎病毒（Infectious laryngotracheitis virus，ILTV），2份鸡传染性支气管炎病毒（IBV）阳性病料进行平行同条件检测，结果均为阴性。表明所建立的RT—PCR特异性好、敏感性高，可用于鸡传染性法氏囊病的临床初步诊断及流行病学调查。

Establishment of a RT-PCR Diagnostic Method for Infectious bursal disease virus

Based on high conservative VP2 gene sequence of IBDV UK661 on GeneBank,a pair of primers were designed and synthesized.The RT-PCR technique for detecting IBDV was established.IBDV-QD RNA was extracted and then was amplified by RT-PCR.The 453bp specific fragments were obtained as positive result.However,the results of detection of ALDV,ILTV,IBV used by RT-PCR in the same condition with IBDV-QD were negative.The amplified sequence of PCR product of IBDV-QD was 97.4%-99.9% in accord with the published data of six IBDV strains on GeneBank.8 shares of positive IBDV materials were detected by RT-PCR and the results were positive,whereas the results of detecting every two shares of positive ALDV,ILTV and IBV materials with the same method were negative.The results concluded that RT-PCR technique in this research had high sensitivity and specificity,and could be applied for IBDV clinical diagnosis.