畜牧兽医学报2012，Vol.43Issue(7)：1096-1102,7.
禽白血病J亚群病毒实时荧光定量PCR检测方法的建立及应用
Development and Application of Real-time PCR for Detection of Avian Leukosis Virus Subgroup J
摘要
Abstract
The organs distributions of Avian Leukosis Virus subgroup J (ALV-J) in chicken were studied through the method of real-time PCR in this article. A SYBR Green I-based real-time PCR was developed for detection of ALV-J by using a pair of primers which was designed to target NX0101 (5 258-5 510 bp). The product of PCR (253 bp) was linked to pMD18-T vector served as standard curve. The sensitivity, specificity and repeatability tests of the method were conducted. The hearts, livers, spleens, lungs, kidneys, thymuses, bursa of Fabricius and glandular stomachs of 20 chickens were detected repeatedly 3 times by this method. The results showed a precise linear relationship with a correlation coefficient of R2 =0. 991 4; the detection limits was 81 copies of DNA plasmid, it was 100 times more sensitive than RT-PCR. The melting curve showed a single peak could only be detected for ALV-J. The variation coefficient of repeatability tests were less than 5%. The copies of ALV-J in thymuses were highest and lungs, spleen, and bursa of Fabricius were higher, and hearts were lowest. Furthermore, the ALV-J copies in natural infection chickens were higher than artificial infection chickens. Real-time PCR about ALV-J gene established in this article was high sensitivity, strong specificity and short testing period. The organs distributions of ALV-J in chicken were preliminarily discussed. The research provided important technical support for the diagnosis and treatment with ALV-J.关键词
禽白血病病毒J亚群/荧光定量PCR/病毒分布Key words
avian leukosis virus subgroup J/ real-time PCR/ virus distribution分类
农业科技引用本文复制引用
董征英,张伟华,李冰,常维山..禽白血病J亚群病毒实时荧光定量PCR检测方法的建立及应用[J].畜牧兽医学报,2012,43(7):1096-1102,7.基金项目
国家公益性行业(农业)科研专项(200803019) （农业）
