关键词: 禽白血病病毒J亚群, 荧光定量PCR, 病毒分布

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 glandular 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.

Key words: avian leukosis virus subgroup J, realtime PCR, virus distribution