PMID- 30145179 OWN - NLM STAT- MEDLINE DCOM- 20181211 LR - 20181211 IS - 1879-0984 (Electronic) IS - 0166-0934 (Linking) VI - 261 DP - 2018 Nov TI - Development and application of a novel Bio-Plex suspension array system for high-throughput multiplexed nucleic acid detection of seven respiratory and reproductive pathogens in swine. PG - 104-111 LID - S0166-0934(18)30154-X [pii] LID - 10.1016/j.jviromet.2018.08.017 [doi] AB - The aim of this study was to develop a multiple PCR assay based on the suspension array system for the simultaneous detection of respiratory and reproductive pathogens in swine. Pseudorabies virus (PRV), Japanese encephalitis virus (JEV), classic swine fever virus (CFSV), African swine fever virus (ASFV), porcine circovirus type 2 (PCV-2), porcine reproductive and respiratory syndrome virus (PRRSV) and porcine parvovirus (PPV) are the major respiratory and reproductive viral pathogens in pig farms. Seven pairs of specific primers and probes were designed, and the multiple PCR was performed, with the PCR products hybridized to beads coupled to probes, which were then detected by Bio-Plex suspension array system. The limit of detection, specificity and repeatability of this method was determined. The assay was further tested using 137 clinical samples, and the results were compared with conventional PCR to evaluate the ability of the method to diagnose porcine viruses. The results showed that the assay had a high degree of specificity and repeatability, and the simultaneous detection limit for the seven viruses reached 10(3) copies/muL. Forty-nine of the clinical samples tested positive for at least one of the viruses, the principal viral infections in the clinical samples were PCV-2 and PRRSV. The suspension method represented a rapid, specific and high-throughput tool for single or mixed detection of the seven porcine viruses simultaneously, and has great significance for the development of liquid chip techniques for the diagnosis of diseases in animals. CI - Copyright (c) 2018 Elsevier B.V. All rights reserved. FAU - Xiao, Lu AU - Xiao L AD - Sichuan Animal Science Academy, Chengdu, 610066, China; Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China. FAU - Wang, Yin AU - Wang Y AD - College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China. Electronic address: wenjiangwangyin@126.com. FAU - Kang, Runmin AU - Kang R AD - Sichuan Animal Science Academy, Chengdu, 610066, China; Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China. FAU - Wu, Xulong AU - Wu X AD - College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China. FAU - Lin, Hua AU - Lin H AD - Sichuan Entry-Exit Inspection and Quarantine Bureau, Chengdu, Sichuan, 610041, China. FAU - Ye, Yonggang AU - Ye Y AD - Sichuan Animal Science Academy, Chengdu, 610066, China; Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China. FAU - Yu, Jifeng AU - Yu J AD - Sichuan Animal Science Academy, Chengdu, 610066, China; Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China. FAU - Ye, Jianqiang AU - Ye J AD - Sichuan Animal Science Academy, Chengdu, 610066, China; Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China. FAU - Xie, Jing AU - Xie J AD - Sichuan Animal Science Academy, Chengdu, 610066, China; Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China. FAU - Cao, Ye AU - Cao Y AD - Sichuan Animal Science Academy, Chengdu, 610066, China; Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China. FAU - Wei, Yong AU - Wei Y AD - Sichuan Animal Science Academy, Chengdu, 610066, China; Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China. FAU - Liao, Dangjin AU - Liao D AD - Sichuan Animal Science Academy, Chengdu, 610066, China; Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China. FAU - Pan, Meng AU - Pan M AD - Sichuan Animal Science Academy, Chengdu, 610066, China; Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China. FAU - Lin, Yi AU - Lin Y AD - Sichuan Animal Science Academy, Chengdu, 610066, China; Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China. FAU - Dai, Zhuojian AU - Dai Z AD - Sichuan Animal Science Academy, Chengdu, 610066, China; Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China. FAU - Li, Xingyu AU - Li X AD - Sichuan Animal Science Academy, Chengdu, 610066, China; Key Laboratory of Animal Genetics and Breeding of Sichuan Province, Chengdu, 611130, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20180824 PL - Netherlands TA - J Virol Methods JT - Journal of virological methods JID - 8005839 RN - 0 (DNA Primers) RN - 0 (Oligonucleotide Probes) SB - IM MH - Animals MH - DNA Primers/genetics MH - High-Throughput Screening Assays/*methods MH - Molecular Diagnostic Techniques/methods MH - Multiplex Polymerase Chain Reaction/*methods MH - Oligonucleotide Probes/genetics MH - Reproducibility of Results MH - Reproductive Tract Infections/diagnosis/*veterinary/virology MH - Respiratory Tract Infections/diagnosis/*veterinary/virology MH - Sensitivity and Specificity MH - Swine MH - Swine Diseases/*diagnosis/virology MH - Time Factors MH - Virus Diseases/diagnosis/*veterinary/virology MH - Viruses/genetics/*isolation & purification OTO - NOTNLM OT - High-throughput OT - Molecular diagnostic OT - Multiplex PCR OT - Suspension array EDAT- 2018/08/27 06:00 MHDA- 2018/12/12 06:00 CRDT- 2018/08/27 06:00 PHST- 2018/03/22 00:00 [received] PHST- 2018/08/17 00:00 [revised] PHST- 2018/08/22 00:00 [accepted] PHST- 2018/08/27 06:00 [pubmed] PHST- 2018/12/12 06:00 [medline] PHST- 2018/08/27 06:00 [entrez] AID - S0166-0934(18)30154-X [pii] AID - 10.1016/j.jviromet.2018.08.017 [doi] PST - ppublish SO - J Virol Methods. 2018 Nov;261:104-111. doi: 10.1016/j.jviromet.2018.08.017. Epub 2018 Aug 24.