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.
出处
《扬州大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
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)
