PMID- 26706731
OWN - NLM
STAT- MEDLINE
DCOM- 20161213
LR  - 20191210
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Linking)
VI  - 232
DP  - 2016 Jun
TI  - Simultaneous typing of seven porcine pathogens by multiplex PCR with a GeXP 
      analyser.
PG  - 21-8
LID - S0166-0934(15)00388-2 [pii]
LID - 10.1016/j.jviromet.2015.12.004 [doi]
AB  - A novel high-throughput method was developed for simultaneous detection and 
      differentiation of seven porcine pathogens by multiplex PCR based on a GenomeLab 
      Gene Expression Profiler (GeXP) analyser. The pathogens included in this study 
      were pseudorabies virus (PRV), classical swine fever virus (CSFV), African swine 
      fever virus (ASFV), porcine reproductive and respiratory syndrome virus (PRRSV), 
      porcine parvovirus (PPV), porcine circovirus type 2 (PCV-2) and Japanese 
      encephalitis virus (JEV). Seven pairs of chimeric primers, consisting of a 
      pathogen-specific sequence fused to a universal sequence at the 5' end, were used 
      to initiate the PCR, after which a set of universal primers was used for the 
      subsequent cycles of the PCR. Amplification products were separated by capillary 
      electrophoresis and identified using fluorescence spectrophotometry. The 
      specificity of the GeXP assay was examined with single and mixed pathogen 
      cDNA/DNA templates. The specific DNA product amplification peaks of seven 
      pathogens were observed on the GeXP analyser. Negative controls did not produce 
      DNA products. The sensitivity was evaluated by performing the assay on serial 
      10-fold dilutions of the plasmids containing the target sequence. Under optimised 
      conditions this assay achieved a sensitivity of 100-1000 copies/muL for a single 
      virus and 1000 copies/muL when all of the seven pre-mixed viral targets were 
      present. Furthermore, the GeXP-PCR assay was 100% specific when 58 clinical 
      samples were tested in comparison with the conventional PCR method. In 
      conclusion, the GeXP assay is a rapid, cost-effective, sensitive, specific and 
      high throughput method for simultaneously detecting seven pathogens that infect 
      swine.
CI  - Copyright (c) 2015 Elsevier B.V. All rights reserved.
FAU - Hu, Ling
AU  - Hu L
AD  - Animal Quarantine Laboratory, College of Veterinary Medicine, Sichuan 
      Agricultural University, Chengdu 611130, Sichuan, China; Zigong Center for Animal 
      Disease Control and Prevention, Zigong 643000, Sichuan, China.
FAU - Lin, Xingyu
AU  - Lin X
AD  - Animal Quarantine Laboratory, College of Veterinary Medicine, Sichuan 
      Agricultural University, Chengdu 611130, Sichuan, China.
FAU - Nie, Fuping
AU  - Nie F
AD  - Chongqing Entry-Exit Inspection and Quarantine Bureau of China, Chongqing 400020, 
      China; Chongqing Import and Export Food Safety Engineering Center, Chongqing 
      400020, China.
FAU - ZexiaoYang
AU  - ZexiaoYang
AD  - Animal Quarantine Laboratory, College of Veterinary Medicine, Sichuan 
      Agricultural University, Chengdu 611130, Sichuan, China.
FAU - Yao, Xueping
AU  - Yao X
AD  - Animal Quarantine Laboratory, College of Veterinary Medicine, Sichuan 
      Agricultural University, Chengdu 611130, Sichuan, China.
FAU - Li, Guili
AU  - Li G
AD  - Animal Quarantine Laboratory, College of Veterinary Medicine, Sichuan 
      Agricultural University, Chengdu 611130, Sichuan, China.
FAU - Wu, Xulong
AU  - Wu X
AD  - Animal Quarantine Laboratory, College of Veterinary Medicine, Sichuan 
      Agricultural University, Chengdu 611130, Sichuan, China.
FAU - Ren, Meishen
AU  - Ren M
AD  - Animal Quarantine Laboratory, College of Veterinary Medicine, Sichuan 
      Agricultural University, Chengdu 611130, Sichuan, China.
FAU - Wang, Yin
AU  - Wang Y
AD  - Animal Quarantine Laboratory, College of Veterinary Medicine, Sichuan 
      Agricultural University, Chengdu 611130, Sichuan, China; Key Laboratory of Animal 
      Disease and Human Health of Sichuan Province, Chengdu 611100, Sichuan, China. 
      Electronic address: yaanwangyin@tom.com.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20151217
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (DNA Primers)
SB  - IM
MH  - Animals
MH  - DNA Primers/genetics
MH  - Electrophoresis, Capillary
MH  - High-Throughput Screening Assays/methods
MH  - Molecular Diagnostic Techniques/*methods
MH  - Multiplex Polymerase Chain Reaction/*methods
MH  - Sensitivity and Specificity
MH  - Spectrophotometry
MH  - Swine
MH  - Swine Diseases/*diagnosis/virology
MH  - Veterinary Medicine/*methods
MH  - Virus Diseases/diagnosis/*veterinary/virology
MH  - Viruses/*classification/genetics/*isolation & purification
OTO - NOTNLM
OT  - GeXP analyser
OT  - Multiplex detection
OT  - Reproductive pathogens
EDAT- 2015/12/27 06:00
MHDA- 2016/12/15 06:00
CRDT- 2015/12/27 06:00
PHST- 2015/06/02 00:00 [received]
PHST- 2015/12/09 00:00 [revised]
PHST- 2015/12/10 00:00 [accepted]
PHST- 2015/12/27 06:00 [entrez]
PHST- 2015/12/27 06:00 [pubmed]
PHST- 2016/12/15 06:00 [medline]
AID - S0166-0934(15)00388-2 [pii]
AID - 10.1016/j.jviromet.2015.12.004 [doi]
PST - ppublish
SO  - J Virol Methods. 2016 Jun;232:21-8. doi: 10.1016/j.jviromet.2015.12.004. Epub 
      2015 Dec 17.