摘要
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。
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.
出处
《动物医学进展》
CSCD
2008年第9期5-8,共4页
Progress In Veterinary Medicine
基金
兽医微生物菌种资源标准化-朊病毒类资源标准化整理整合及共享项目科技部(2005KDA21205-7)
