PMID- 22575688
OWN - NLM
STAT- MEDLINE
DCOM- 20120830
LR  - 20120511
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Linking)
VI  - 183
IP  - 1
DP  - 2012 Jul
TI  - Development of multiplex PCR for simultaneous detection of six swine DNA and RNA 
      viruses.
PG  - 69-74
LID - 10.1016/j.jviromet.2012.03.034 [doi]
AB  - Uniplex and multiplex reverse transcription-polymerase chain reaction (RT-PCR) 
      and PCR protocols were developed and evaluated subsequently for its effectiveness 
      in detecting simultaneously single and mixed infections in swine. Specific 
      primers for three DNA viruses and three RNA viruses, including classical swine 
      fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), 
      Japanese encephalitis virus (JEV), porcine circovirus type 2 (PCV2), porcine 
      pseudorabies virus (PRV) and porcine parvovirus (PPV) were used for testing 
      procedure. A single nucleic acid extraction protocol was adopted for the 
      simultaneous extraction of both RNA and DNA viruses. The multiplex PCR consisted 
      with two-step procedure which included reverse transcription of RNA virus and 
      multiplex PCR of viral cDNA and DNA. The multiplex PCR assay was shown to be 
      sensitive detecting at least 450pg of viral genomic DNA or RNA from a mixture of 
      six viruses in a reaction. The assay was also highly specific in detecting one or 
      more of the same viruses in various combinations in specimens. Thirty clinical 
      samples and aborted fetuses collected from 4- to 12-week-old piglets were 
      detected among 39 samples tested by both uniplex and multiplex PCR, showing 
      highly identification. Because of the sensitivity and specificity, the multiplex 
      PCR is a useful approach for clinical diagnosis of mixed infections of DNA and 
      RNA viruses in swine.
CI  - Copyright (c) 2012 Elsevier B.V. All rights reserved.
FAU - Xu, Xin-Gang
AU  - Xu XG
AD  - College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 
      712100, China.
FAU - Chen, Guang-Da
AU  - Chen GD
FAU - Huang, Yong
AU  - Huang Y
FAU - Ding, Li
AU  - Ding L
FAU - Li, Zhao-Cai
AU  - Li ZC
FAU - Chang, Ching-Dong
AU  - Chang CD
FAU - Wang, Chi-Young
AU  - Wang CY
FAU - Tong, De-Wen
AU  - Tong DW
FAU - Liu, Hung-Jen
AU  - Liu HJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120403
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (DNA Primers)
SB  - IM
MH  - Animals
MH  - Coinfection/diagnosis/veterinary/virology
MH  - DNA Primers/genetics
MH  - DNA Virus Infections/diagnosis/*veterinary/virology
MH  - DNA Viruses/genetics/*isolation & purification
MH  - Multiplex Polymerase Chain Reaction/*methods
MH  - RNA Virus Infections/diagnosis/*veterinary/virology
MH  - RNA Viruses/genetics/*isolation & purification
MH  - Sensitivity and Specificity
MH  - Swine
MH  - Swine Diseases/*diagnosis/*virology
MH  - Veterinary Medicine/methods
MH  - Virology/methods
EDAT- 2012/05/12 06:00
MHDA- 2012/08/31 06:00
CRDT- 2012/05/12 06:00
PHST- 2011/08/26 00:00 [received]
PHST- 2012/03/21 00:00 [revised]
PHST- 2012/03/26 00:00 [accepted]
PHST- 2012/05/12 06:00 [entrez]
PHST- 2012/05/12 06:00 [pubmed]
PHST- 2012/08/31 06:00 [medline]
AID - S0166-0934(12)00113-9 [pii]
AID - 10.1016/j.jviromet.2012.03.034 [doi]
PST - ppublish
SO  - J Virol Methods. 2012 Jul;183(1):69-74. doi: 10.1016/j.jviromet.2012.03.034. Epub 
      2012 Apr 3.