›› 2019, Vol. 46 ›› Issue (5): 1532-1540.doi: 10.16431/j.cnki.1671-7236.2019.05.033

• Clinical Veterinary Medicine • Previous Articles     Next Articles

Research on Multiplex PCR Combined with DNA Microarray for Detecting Five Kinds of Reproductive Disease Virus

LIU Shengli1, LIU Lingling2, LYU Yujin2, WU Fengsun2, LI Wengang2   

  1. 1. College of Livestock Husbandry and Veterinary Engineering, Henan Agricultural University, Zhengzhou 450002, China;
    2. College of Veterinary Medicine, Henan University of Auimal Husbandry Economy, Zhengzhou 450046, China
  • Received:2018-10-08 Online:2019-05-20 Published:2019-05-20

Abstract:

A multiplex PCR combined with DNA microarray method was developed for detection of 5 swine reproductive diseases virus in this study.Five pairs of specific primers and probes were designed according to the published genomic sequences of swine fever virus (CSFV),porcine parvovirus (PPV),porcine reproductive and respiratory syndrome virus (PRRSV),Japanese encephalitis virus (JEV) and porcine circovirus type 2 (PCV2).The corresponding oligonucleotide chips were prepared,16 clinical samples and the standard strains of five reproductive viruses were tested.Specific gene fragments with fluorescent label were amplified by multiplex PCR and hybridized with microarray that immobilized with specific probes.The results showed that the multiplex PCR combined with DNA microarray to detect viral diseases was highly specific and stable,and the sensitivity was up to 102 copies/μL.The test results of 16 clinical samples showed a positive rate of 87.5% (14/16).These data indicated that the sensitive and specific detection method could be applied in these 5 viruses detection for clinical diagnosis and pathogenic epidemiologic investigation.

Key words: multiplex PCR; microarray; CSFV; PPV; PRRSV; JEV; PCV2

CLC Number: