›› 2012, Vol. 39 ›› Issue (10): 37-41.

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Development of Gene Chip Method for Detecting AHSV,WNV and ECV

ZHAO Yin-ze1,2, WANG Lin1,2, XING You-shang1,2, ZHANG Can3   

  1. 1. Shenyang Agricultural University,Shenyang 110866,China;
    2. Beijing Entry-exit Inspection and Quarantine Bureau,Beijing 100026,China;
    3. Institute of Hygiene and Environmental Medicine, Academy of Military Medical Sciences,Tianjin 300050, China
  • Received:2012-02-22 Online:2012-10-20 Published:2012-10-19

Abstract: In order to establish a gene-chip detection method of equine virus, the highly conserved DNAs of African horse sickness virus (AHSV ) was obtained by artificial splicing, West Nile virus(WNV) and equine corona virus(ECV) were acquired by molecular cloning, and then spotted on the treated glass slides as the diagnostic gene-chip. And the cDNAs amplified by multiple asymmetric PCR with the template of splicing and cloned nucleic acid sequence were labeled with fluorescence as probes. Following specific hybridization of deposited gene chip and labeled probes, fluorescence signals were scanned by laser scanner and the obtained image was analyzed by Qiamt Array software with the digital computer. The results showed that the prepared gene chip could detect and distinguish the three equine viruses. And its sensitivity was about 104 copies of AHSV and 102 of ECV and WNV. The hybridization specificity was confirmed by the presence of fluorescence signals on the corresponding sites with samples from the three relevant viruses DNA template and by the absence of positive signals with the specimens from irrelevant viruses was negative by gene chip. The evidence suggested that gene chip, which was quick, specific, sensitive, and reliable, could provide a practical alternative to screen and quarantine a large number of samples within a very short period of time.

Key words: African horse sickness virus; West Nile virus; equine corona virus; gene chip; detection

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