ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2018, Vol. 49 ›› Issue (6): 1314-1319.doi: 10.11843/j.issn.0366-6964.2018.06.025

Previous Articles     Next Articles

Establishment and Application of LAMP Method for Detection of Porcine Circovirus Type 3

JIANG Chen-long1, YAN Xiu-wen1, ZHANG Ri-teng2, LI Yong2, JIANG Ping2, BAI Juan2*   

  1. 1. Key Laboratory of Zoology of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China;
    2. Key Laboratory of Animal Bacteriology of Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
  • Received:2017-10-20 Online:2018-06-23 Published:2018-06-22

Abstract:

In order to diagnose and detect porcine circovirus type 3 (PCV3), in this research, specific primers were designed according to the conserved sequence of ORF2 gene. By optimizing the concentration of Bst DNA polymerase, the ratio of primers, Mg2+, dNTPs, betaine and reaction condition, the loop-mediated isothermal nucleic acid amplification (LAMP) was established. The results manifested that ladder strips were observed after 36min with a constant temperature, and the product could be judged by SYBR Ⅰ staining. The detection limit of this way was 1.0×101 copies·μL-1 which had no cross reaction with porcine reproductive and respiratory syndrome virus (PRRSV), porcine pseudorabies virus (PRV), classical swine fever virus (CSFV), porcine circovirus type 1 (PCV1) and type 2 (PCV2). From 68 lymph nodes of diseased pigs in respiratory system, the positive rate of 30.9% tested by LAMP had a high coincidence rate of 95.6% (65/68) with that by PCR detection. Compared with traditional technology, this method is more sensitive, simpler and can be used for detection and clinical diagnosis of PCV3.

Key words: PCV3, RT-LAMP, detection

CLC Number: