ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2012, Vol. 43 ›› Issue (7): 1096-1102.doi:

• 预防兽医 • Previous Articles     Next Articles

Development and Application of Realtime PCR for Detection of Avian Leukosis Virus Subgroup J

DONG Zhengying1, ZHANG Weihua2, LI Bing1, CHANG Weishan1*   

  1. 1. Cellular Immunology Laboratory, Preventive Veterinary Medicine of College of Animal Science and Technology, Shandong Agricultural University, Taian 271018, China; 2. Shandong Huamutianyuan A & H Stock Co. Ltd, Jinan 250100, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2012-07-27 Published:2012-07-27
  • Contact: CHANG Weishan1

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.