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BioMed Research International
Volume 2017, Article ID 7243909, 8 pages
Research Article

A Novel Capillary Electrophoresis-Based High-Throughput Multiplex Polymerase Chain Reaction System for the Simultaneous Detection of Nine Pathogens in Swine

1College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China
2Inspection and Quarantine Technical Center, Sichuan Entry-Exit Inspection and Quarantine Bureau, Chengdu 610041, China
3Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu 611130, China
4College of Life Science, Sichuan Agricultural University, Ya’an 625014, China

Correspondence should be addressed to Yin Wang; moc.361@niygnawnaay

Received 31 March 2017; Accepted 14 May 2017; Published 11 June 2017

Academic Editor: Yanjin Zhang

Copyright © 2017 Xu-long Wu et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Here we aimed to develop a capillary electrophoresis-based high-throughput multiplex polymerase chain reaction (PCR) system for the simultaneous detection of nine pathogens in swine. Nine pairs of specific primers and a set of universal primers were designed; the multiplex PCR was established. The specificity and cross-reactivity of this assay were examined, and the detection limit was determined using serial 10-fold dilutions of plasmids containing the target sequences. The assay was further tested using 144 clinical samples. We found that the nine specific amplification peaks were observed, and the assay had a high degree of specificity, without nonspecific amplification. The simultaneous detection limit for the nine viruses reached 10000 copies μL−1 when all of the premixed viral targets were present. Seventy-seven of the clinical samples tested positive for at least one of the viruses; the principal viral infections in the clinical samples were porcine circovirus type 2 and porcine reproductive and respiratory syndrome virus. This approach has much potential for further development of high-throughput detection tools for the diagnosis of diseases in animals.