PMID- 30951787
OWN - NLM
STAT- MEDLINE
DCOM- 20200603
LR  - 20200603
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Print)
IS  - 0166-0934 (Linking)
VI  - 269
DP  - 2019 Jul
TI  - A multiplex RT-PCR assay for rapid and simultaneous detection of four RNA viruses 
      in swine.
PG  - 38-42
LID - S0166-0934(19)30024-2 [pii]
LID - 10.1016/j.jviromet.2019.04.001 [doi]
AB  - A multiplex reverse transcription polymerase chain rection (mRT-PCR) was 
      developed for simultaneous detection of four RNA viruses in swine. The conserved 
      target sequences directed to classical swine fever virus (CSFV), porcine 
      reproductive and respiratory syndrome virus (PRRSV), porcine epidemic diarrhea 
      virus (PEDV) and transmissible gastroenteritis coronavirus (TGEV) were selected 
      based on alignments of genomic sequences and then specific primers were designed. 
      The mRT-PCR assay was developed and evaluated for its specificity and 
      sensitivity. The expected product from the single viral template was amplified by 
      mRT-PCR and no spurious PCR amplification occurred from the genomic RNA or DNA of 
      other pathogens. For single virus or different combinations of two viruses the 
      detection limit of mRT-PCR was consistent with a single RT-PCR wtith 1 x 10(3) 
      copies. For different combinations of the three viruses or four viruses, 
      sensitivity of PEDV detection partially decreased. All of positive clinical 
      specimens by the mRT-PCR were identically confirmed using Taqman RT-qPCR. 
      Therefore, the mRT-PCR is a useful tool for epidemiological studies and 
      laboratory diagnosis of single virus and/or mixed infections in swine.
CI  - Copyright (c) 2019 Elsevier B.V. All rights reserved.
FAU - Zhao, Yan
AU  - Zhao Y
AD  - State Key Laboratory of Virology, Modern Virology Research Center, College of 
      Life Sciences, Wuhan University, Wuhan 430072, China.
FAU - Liu, Feifei
AU  - Liu F
AD  - State Key Laboratory of Virology, Modern Virology Research Center, College of 
      Life Sciences, Wuhan University, Wuhan 430072, China.
FAU - Li, Qingmei
AU  - Li Q
AD  - Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, 
      Zhengzhou 450002, China.
FAU - Wu, Mengfan
AU  - Wu M
AD  - State Key Laboratory of Virology, Modern Virology Research Center, College of 
      Life Sciences, Wuhan University, Wuhan 430072, China.
FAU - Lei, Lei
AU  - Lei L
AD  - State Key Laboratory of Virology, Modern Virology Research Center, College of 
      Life Sciences, Wuhan University, Wuhan 430072, China.
FAU - Pan, Zishu
AU  - Pan Z
AD  - State Key Laboratory of Virology, Modern Virology Research Center, College of 
      Life Sciences, Wuhan University, Wuhan 430072, China. Electronic address: 
      zspan@whu.edu.cn.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190402
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (DNA Primers)
RN  - 0 (RNA, Viral)
SB  - IM
MH  - Animals
MH  - Classical Swine Fever Virus/isolation & purification
MH  - DNA Primers/genetics
MH  - Multiplex Polymerase Chain Reaction/methods/*veterinary
MH  - Porcine epidemic diarrhea virus/isolation & purification
MH  - Porcine respiratory and reproductive syndrome virus/isolation & purification
MH  - RNA Virus Infections/*veterinary/virology
MH  - RNA Viruses/classification/*isolation & purification
MH  - RNA, Viral/genetics
MH  - Sensitivity and Specificity
MH  - Swine
MH  - Swine Diseases/*diagnosis/*virology
MH  - Transmissible gastroenteritis virus/isolation & purification
PMC - PMC7113762
OTO - NOTNLM
OT  - Multiplex RT-PCR
OT  - Simultaneous detection
OT  - Swine
OT  - Virus infection
EDAT- 2019/04/06 06:00
MHDA- 2020/06/04 06:00
PMCR- 2019/04/02
CRDT- 2019/04/06 06:00
PHST- 2019/01/15 00:00 [received]
PHST- 2019/03/30 00:00 [revised]
PHST- 2019/04/01 00:00 [accepted]
PHST- 2019/04/06 06:00 [pubmed]
PHST- 2020/06/04 06:00 [medline]
PHST- 2019/04/06 06:00 [entrez]
PHST- 2019/04/02 00:00 [pmc-release]
AID - S0166-0934(19)30024-2 [pii]
AID - 10.1016/j.jviromet.2019.04.001 [doi]
PST - ppublish
SO  - J Virol Methods. 2019 Jul;269:38-42. doi: 10.1016/j.jviromet.2019.04.001. Epub 
      2019 Apr 2.