鸡传染性支气管炎病毒的快速检测  

PCR Method for the Rapid Detection of IBV

作　　者：刘思国[1] 康丽娟[2] 江国托 刘明春[4] 孔宪刚[1] 杨宏伟 刘忠贵[6] 李从寿[7] 宋春青 霍兰享 卢景良[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001 [2]大连市畜牧兽医技术服务中心 [3]大连三仪生物工程研究所 [4]沈阳农业大学畜牧兽医学院 [5]吉林省畜禽总公司 [6]东北农业大学动物医学院 [7]四川省西南民族学院 [8]天津市西青区王稳庄镇兽医站 [9]天津市西青区南河镇畜牧兽医站

出　　处：《中国预防兽医学报》2002年第5期386-390,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家攀登计划B类项目"85_44_0 1_44"资助课题

摘　　要：本试验建立了一种快速检测鸡传染性支气管炎病毒的通用性PCR方法。通过对纯化后IBV核蛋白基因进行的直接PCR及套式PCR扩增表明 ,两次PCR产物的大小为 0 .7kb和 0 .5kb ,与实际设计相符。而对照的NDV及IBDV的PCR扩增产物均为阴性。用IBVHB(华北 )株感染SPF鸡后 ,应用我们所建立的PCR方法去检测不同时期所采集的肾脏、气管、盲肠扁桃体、肺脏和肝脏中的IBV ,在SPF鸡感染IBV后的 2 1天仍然能够检测到IBV的基因组 ,Southernblot杂交试验证明了我们获得的PCR产物为IBV的核蛋白基因。对 30份自然感染病料的检测表明 ,本实验所建立的PCR方法检测阳性数为 15份 ,阳性率为 5 0 % (15 /30 ) ;鸡胚接种检测的阳性数为 17份 ,阳性率为 5 6 .7% (17/30 ) ;IFA检测的阳性数为 11份 ,阳性率为 36 .7% (11/30 )。PVR检测法与鸡胚接种方法的阳性符合率为 93.3% (14/15 ) ,统计学分析表明 ,P >0 .0 5 ,差异不显著 ,而PCR检测法与IFA的阳性符合率为 6 0 % (9/15 )。结果证明本实验所建立的PCR检测方法具有敏感、特异、快速等特点 。A PCR method was developed for the rapid detection of IBV.Direct and nested PCR for amplifying purified N gene showed that two products were 0.7kb and 0.5kb,which were identical to initial designed, while the PCR products of controlled NDV and IBDV were negative. SPF chickens were infected with IBV HB strain and their kidney,trachea,lung and liver were collected at different time and detected by using our PCR method.The result showed that IBV was still detected at 21 st day post infection. Southern blotting hybridization proved that our PCR product was IBV N gene.30 samples tested by PCR, chickens embryo culture and IFA.The positive nubmers wers 15,17 and 11,respectively. The PCR and chickens embryo culture test shared the same results which was up to 93.3%(14/15)and there were no differentiation(P>0.05)between the two methods.Our PCR (method) is more sensitive,specific,rapid than IFA and is competent of the detection for different IBV strains.

关 键 词：鸡 传染性 支气管炎 病毒 快速检测 

分 类 号：S858.31[农业科学—临床兽医学]