10种猪常见病毒基因芯片检测方法的建立及初步应用

doi:10.11843/j.issn.0366-6964.2018.10.021

Abstract

Abstract:

To develop a new reliable method for initial diagnosis and epidemiological investigation of ten common diseases of pig, an DNA microarray was established for simultaneous detection of porcine reproductive and respiratory syndrome virus(PRRSV), classical swine fever virus (CSFV), pseudorabies virus (PRV), porcine circovirus 2 (PCV2), porcine parvovirus (PPV), porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis of swine (TGEV), swine influenza virus (SIV), porcine cytomegalovirus (PCMV) and food-and-mouth disease virus (FMDV). According to the similarity of the ten viral sequences in the GenBank database, ten pairs of specific primers were designed and ten kinds of virus-specific conservative regions were amplified by PCR, respectively. Then the purified target genes were fixed on the amino group modified slides. The probes labeled with Cy3 were amplified from samples by PCR and hybridized with the cDNA array on the slides. The hybridized results were scanned by scanner. The gene chip detection method was established and its specificity, sensitivity and repeatability were detected. The coincidence test between cDNA chip and PCR/RT-PCR was performed in 1 006 samples. The results showed that this cDNA array was very specific and sensitive, the minimum sensitivity was 10⁵ copies·μL^-1. The coincidence rate between cDNA chip and PCR/RT-PCR was 96.79%. In all, the gene chip detection technology for ten kinds of common swine pathogenic viruses was successfully developed and can be applied to swine pathogens detection in clinic.

CLC Number:

S858.285.3

LUO Ning, WANG Qun, ZHANG Xiao-long, FAN Bao-hai, JIANG Fan, SHI Ling-ling, WANG Dong-dong, YANG Zong-tong, WANG Shou-chun, XU Biao, YIN Yan-bo. Research on DNA Microarray for Detecting Ten Different Virus from Swine[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(10): 2249-2260.

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References

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Research on DNA Microarray for Detecting Ten Different Virus from Swine

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