J亚群禽白血病病毒TD-PCR检测方法的建立及初步应用  

Establishment and application of TD-PCR for detection of subgroup J avian leukosis virus

作　　者：杭柏林[1,2] 秦爱建[1] 钱琨[1] 金文杰[1] 沈海玉[1] 吴晓平[1] 仇钰[1] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]河南科技学院动物科学学院,河南新乡453003

出　　处：《扬州大学学报（农业与生命科学版）》2010年第4期40-44,共5页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家公益性行业科研专项基金资助项目(200803019);国家自然科学基金联合基金重点项目(U0831002);江苏省高校自然科学重大基础研究项目(07KJA23021)

摘　　要：采用J亚群禽白血病病毒(ALV-J)感染DF1细胞提取的总DNA作为模板,以H5和H7为特异性引物,建立检测ALV-J的降落PCR(TD-PCR)方法,并对反应体系进行优化。结果表明:建立TD-PCR方法灵敏度高、特异性强,可检测0.105 ng的DNA,该方法与网状内皮组织增生症病毒、马立克病病毒、鸡贫血病毒及I群禽腺病毒没有交叉反应。运用TD-PCR、ELISA以及病毒分离对临床病料进行检测,TD-PCR方法与ELISA检测结果一致(100%),均比病毒分离检出率(75%)高。证明TD-PCR检测ALV-J的方法具有快速、灵敏、特异等优点,可满足临床、生产实际检测的需要。In order to establish touchdown polymerase chain reaction（TD-PCR） tests for detection of subgroup J avian leukosis virus（ALV-J）,we optimized the PCR reaction with the DNA template extracted from ALV-J infected DF1 cells and special primers pair H5/H7.The results showed that the sensitivity of the TD-PCR test was as low as 0.105 ng DNA of sample.It had no cross-reactions with the DNA from reticuloendotheliosis virus,Marek＇s disease virus（CVI988）,chicken anemia virus and subtype I avian adenovirus.The detection rates of TD-PCR for clinical samples were the same as that of ELISA（100%）,and higher than that of viral isolation（75%）.All these data suggest that the established TD-PCR test is a rapid,sensitive and special method for detection of ALV-J,which is good enough to be used in the field and lab research.

关 键 词：J亚群禽白血病病毒 降落PCR 病毒检测 

分 类 号：S852.65[农业科学—基础兽医学]