PMID- 32304349 OWN - NLM STAT- MEDLINE DCOM- 20210402 LR - 20210402 IS - 1865-1682 (Electronic) IS - 1865-1674 (Linking) VI - 67 IP - 6 DP - 2020 Nov TI - Simultaneous identification of 6 pathogens causing porcine reproductive failure by using multiplex ligation-dependent probe amplification. PG - 2467-2474 LID - 10.1111/tbed.13585 [doi] AB - We developed a multiplex ligation-dependent probe amplification (MLPA) assay for the simultaneous detection of 6 clinically relevant viral pathogens causing porcine reproductive failure, that is porcine reproductive and respiratory syndrome virus (PRRSV), Japanese encephalitis virus (JEV), classical swine fever virus (CSFV), porcine circovirus type 2 (PCV2), pseudorabies virus (PRV) and porcine parvovirus (PPV). The limits of detection for the assay varied among the 6 target organisms from 1 to 8 copies per MLPA assay. The MLPA assay was evaluated with 346 heparinized porcine umbilical cord blood specimens, and the results of the assay were compared to those of real-time PCR. The MLPA assay showed specificities and sensitivities of 99.2% and 100%, respectively, for PRRSV; 100% and 100%, respectively, for CSFV, PCV2, PRV and PPV. No sample was found to be positive for JEV by either the MLPA assay or the real-time PCR. In conclusion, the MLPA assay has comparable clinical sensitivity to that of real-time PCR assay and provides a useful tool for fast screening porcine reproductive failure-associated viruses. CI - (c) 2020 Blackwell Verlag GmbH. FAU - Zhou, Yingshan AU - Zhou Y AUID- ORCID: 0000-0002-8011-431X AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. FAU - Chen, Lin AU - Chen L AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. FAU - Zhang, Lifei AU - Zhang L AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. FAU - Shao, Chunyan AU - Shao C AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. FAU - Sun, Jing AU - Sun J AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. FAU - Jiang, Sheng AU - Jiang S AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. FAU - Song, Quanjiang AU - Song Q AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. FAU - Zhou, Bin AU - Zhou B AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. FAU - Yang, Yongchun AU - Yang Y AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. FAU - Dong, Wanyu AU - Dong W AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. FAU - Yang, Yang AU - Yang Y AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. FAU - Wei, Fangfang AU - Wei F AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. FAU - Fang, Weihuan AU - Fang W AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. AD - Zhejiang Provincial Key Lab of Preventive Veterinary Medicine, Institute of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang University, Hangzhou, Zhejiang, China. FAU - Wang, Xiaodu AU - Wang X AUID- ORCID: 0000-0002-0512-9212 AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. FAU - Song, Houhui AU - Song H AUID- ORCID: 0000-0001-6530-5794 AD - Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Zhejiang Provincial Engineering Laboratory for Animal Health Inspection and Internet Technology, College of Animal Science and Technology, College of Veterinary Medicine, Zhejiang A&F University, Lin'an, Zhejiang, China. LA - eng GR - 2017YFC1200500/National Key Research and Development Program of China/ GR - 2019C02043/Key Research and Development Program of Zhejiang Province/ GR - 2019C02052/Key Research and Development Program of Zhejiang Province/ GR - 2018C02028/Key Research and Development Program of Zhejiang Province/ GR - 31902249/National Natural Science Foundation of China/ GR - 31972656/National Natural Science Foundation of China/ GR - 31502127/National Natural Science Foundation of China/ GR - 31602062/National Natural Science Foundation of China/ GR - 31802258/National Natural Science Foundation of China/ GR - 31602119/National Natural Science Foundation of China/ GR - 31502034/National Natural Science Foundation of China/ GR - LQ19C180003/Zhejiang Provincial Natural Science Foundation/ GR - LQ15C180002/Zhejiang Provincial Natural Science Foundation/ GR - LGN18C180001/Zhejiang Provincial Natural Science Foundation/ GR - LQ19C180002/Zhejiang Provincial Natural Science Foundation/ GR - 2013FK023/Zhejiang A&F University/ GR - 2013FR076/Zhejiang A&F University/ GR - 2015FR042/Zhejiang A&F University/ GR - 2018FR009/Zhejiang A&F University/ GR - 2018FR015/Zhejiang A&F University/ GR - Y201533341/Department of Education of Zhejiang Province/ GR - 201810341022/National Students' platform for innovation and entrepreneurship training program/ PT - Journal Article DEP - 20200505 PL - Germany TA - Transbound Emerg Dis JT - Transboundary and emerging diseases JID - 101319538 SB - IM MH - Animals MH - Circovirus/genetics/*isolation & purification MH - Classical Swine Fever Virus/genetics/*isolation & purification MH - Female MH - Fetal Blood/virology MH - Herpesvirus 1, Suid/genetics/*isolation & purification MH - Limit of Detection MH - Multiplex Polymerase Chain Reaction/veterinary MH - Parvovirus, Porcine/genetics/*isolation & purification MH - Porcine respiratory and reproductive syndrome virus/genetics/*isolation & purification MH - Sensitivity and Specificity MH - Swine MH - Swine Diseases/*virology MH - Virus Diseases/*veterinary/virology OTO - NOTNLM OT - Multiplex Ligation-dependent Probe Amplification (MLPA) OT - porcine reproductive failure OT - viral disease EDAT- 2020/04/19 06:00 MHDA- 2021/04/07 06:00 CRDT- 2020/04/19 06:00 PHST- 2019/11/04 00:00 [received] PHST- 2020/03/07 00:00 [revised] PHST- 2020/04/11 00:00 [accepted] PHST- 2020/04/19 06:00 [pubmed] PHST- 2021/04/07 06:00 [medline] PHST- 2020/04/19 06:00 [entrez] AID - 10.1111/tbed.13585 [doi] PST - ppublish SO - Transbound Emerg Dis. 2020 Nov;67(6):2467-2474. doi: 10.1111/tbed.13585. Epub 2020 May 5.