PMID- 25102430
OWN - NLM
STAT- MEDLINE
DCOM- 20150527
LR  - 20191210
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Linking)
VI  - 208
DP  - 2014 Nov
TI  - Development of an EvaGreen-based multiplex real-time PCR assay with melting curve 
      analysis for simultaneous detection and differentiation of six viral pathogens of 
      porcine reproductive and respiratory disorder.
PG  - 56-62
LID - S0166-0934(14)00293-6 [pii]
LID - 10.1016/j.jviromet.2014.06.027 [doi]
AB  - Concurrent infection of pigs with two or more pathogens is common in pigs under 
      intensive rearing conditions. Porcine circovirus type 2 (PCV2), porcine 
      parvovirus (PPV), porcine reproductive and respiratory syndrome virus (PRRSV), 
      classical swine fever virus (CSFV), Japanese encephalitis virus (JEV) and 
      pseudorabies virus (PRV) are all associated with reproductive or respiratory 
      disorders or both and can cause significant economic losses in pig production 
      worldwide. An EvaGreen-based multiplex real-time PCR (EG-mPCR) with melting curve 
      analysis was developed in this study for simultaneous detection and 
      differentiation of these six viruses in pigs. This method is able to detect and 
      distinguish PCV2, PPV, PRRSV, CSFV, JEV and PRV with the limits of detection 
      ranging from 100 to 500 copies/muL, high reproducibility, and intra-assay and 
      inter-assay variation ranging from 0.11 to 3.20%. After validation, a total of 
      118 field samples were tested by the newly developed EG-mPCR. PCV2 was identified 
      in 23%, PPV in 15%, PRRSV in 17% and PRV in 5% of the samples. Concurrent PCV2 
      and PRRSV infection was detected in 6.7%, PCV2 and PPV in 5% and PPV2 and PRRSV 
      infection was detected in 5% of the cases. The agreement of the EG-mPCR and 
      conventional PCR tests was 99.2%. This EG-mPCR will be a useful, rapid, reliable 
      and cost-effective alternative for routine surveillance testing of viral 
      infections in pigs.
CI  - Copyright (c) 2014 Elsevier B.V. All rights reserved.
FAU - Rao, Pinbin
AU  - Rao P
AD  - College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, China.
FAU - Wu, Haigang
AU  - Wu H
AD  - College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, China.
FAU - Jiang, Yonghou
AU  - Jiang Y
AD  - College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, China. 
      Electronic address: yonghoujiang@zstu.edu.cn.
FAU - Opriessnig, Tanja
AU  - Opriessnig T
AD  - The Roslin Institute and The Royal (Dick) School of Veterinary Studies, 
      University of Edinburgh, Midlothian EH25 9RG, UK; Department of Veterinary 
      Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa 
      State University, Ames, IA, USA.
FAU - Zheng, Xiaowen
AU  - Zheng X
AD  - College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, China.
FAU - Mo, Yecheng
AU  - Mo Y
AD  - College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, China.
FAU - Yang, Zongqi
AU  - Yang Z
AD  - College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, China.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140804
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
SB  - IM
MH  - Animals
MH  - Costs and Cost Analysis
MH  - DNA Viruses/classification/genetics/isolation & purification
MH  - Molecular Diagnostic Techniques/economics/methods
MH  - Multiplex Polymerase Chain Reaction/economics/*methods
MH  - RNA Viruses/classification/genetics/isolation & purification
MH  - Real-Time Polymerase Chain Reaction/economics/*methods
MH  - Reproducibility of Results
MH  - Reproductive Tract Infections/diagnosis/*veterinary/virology
MH  - Respiratory Tract Infections/diagnosis/*veterinary/virology
MH  - Sensitivity and Specificity
MH  - Staining and Labeling/methods
MH  - Swine
MH  - Swine Diseases/*diagnosis/virology
MH  - Time Factors
MH  - Transition Temperature
MH  - Veterinary Medicine/economics/*methods
MH  - Virus Diseases/diagnosis/*veterinary/virology
OTO - NOTNLM
OT  - EvaGreen
OT  - Melting curve
OT  - Multiplex real-time PCR
OT  - Pig viruses
OT  - Surveillance
EDAT- 2014/08/08 06:00
MHDA- 2015/05/28 06:00
CRDT- 2014/08/08 06:00
PHST- 2014/02/18 00:00 [received]
PHST- 2014/05/17 00:00 [revised]
PHST- 2014/06/20 00:00 [accepted]
PHST- 2014/08/08 06:00 [entrez]
PHST- 2014/08/08 06:00 [pubmed]
PHST- 2015/05/28 06:00 [medline]
AID - S0166-0934(14)00293-6 [pii]
AID - 10.1016/j.jviromet.2014.06.027 [doi]
PST - ppublish
SO  - J Virol Methods. 2014 Nov;208:56-62. doi: 10.1016/j.jviromet.2014.06.027. Epub 
      2014 Aug 4.