ACTA VETERINARIA ET ZOOTECHNICA SINICA

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Establishment of a Diplex xMAP Array for Detection of PPV and PCV2

WANG Jing-yu1, ZHU Xiang-bo1,2, LIU Ye-bing2*, HAN Xue-qing3*, WANG Hui-yu3
ZHANG Li3, ZHANG Lei2, ZHANG Xiao-jie2, TANG Pan1   

  1. 1. Northwest A & F University, Yangling 712100, China; 2. China Institute of Veterinary Drug
    Control, Beijing 100081, China; 3. Chinese Academy of Inspection and Quarantine,
    Beijing 100029, China
  • Received:2012-03-08 Online:2012-10-25 Published:2012-10-25

Abstract: The aim of this study was to establish a diplex xMAP array, which can detect porcine parvovirus (PPV) and porcine circovius type 2 (PCV2) simultaneously, to meet the requirements of the immigration quarantine and clinical high-throughput detection. According to the structural protein VP2 gene of PPV and PCV2 genome sequences in GenBank, specific probe, primers and probe’s reverse complement sequence of PPV and PCV2 were designed after multiple sequence alignment analysis. The probe were coupled to microspheres, and the reverse primers, probe’s reverse complement sequence were labeled with biotin. The target sequences were amplified by diplex asymmetry PCR, then products of PCR were hybridized with the probes which coupled verify successful, hybridization results were detected by the xMAP instrument, then the diplex xMAP array, which was established through optimizing the reaction conditions, were used to detect clinical samples. The diplex xMAP array, for detecting PPV and PCV2, was established and had a good specificity and sensitivity, and limit of detection were 1.53×104 and 2.58×102 in gene copy respectively. The detection results of 56 clinical samples showed that this method was 100% consistent with PCR. This study established the diplex xMAP array for detecting PPV and PCV2 successfully; the methods could be used for the immigration quarantine and veterinary clinical detection.

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