PMID- 17975735
OWN - NLM
STAT- MEDLINE
DCOM- 20080403
LR  - 20211020
IS  - 0165-7380 (Print)
IS  - 0165-7380 (Linking)
VI  - 32
IP  - 3
DP  - 2008 Mar
TI  - Development of a novel hot-start multiplex PCR for simultaneous detection of 
      classical swine fever virus, African swine fever virus, porcine circovirus type 
      2, porcine reproductive and respiratory syndrome virus and porcine parvovirus.
PG  - 255-62
AB  - A novel hot-start multiplex PCR (mPCR) assay was developed and subsequently 
      evaluated for its effectiveness in simultaneously detecting multiple viral 
      infections of swine. Specific primers for each of five virus genomes, namely 
      classical swine fever virus (CSFV), African swine fever virus (ASFV), porcine 
      circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus 
      (PRRSV) and porcine parvovirus (PPV) were used. Combined nucleic acid 
      purification was carried out using a commercial RNA/DNA extraction kit. The mPCR 
      consisted of a two-step procedure which included reverse transcription and PCR 
      amplification. This mPCR and the corresponding separate assays were evaluated 
      comparatively on serial ten-fold dilutions of each virus. Analysis of the 
      sensitivity in comparison to the corresponding single PCR (sPCR) for the 
      detection of each of the five targets was identical for CSFV, PCV2 and PPV, 1 log 
      lower for PRRSV and 2 logs lower for ASFV. No spurious PCR amplification 
      reactions among all five pathogens were noticed with various amounts of DNA and 
      RNA mixtures. All the uninfected controls were scored negative. The relative 
      efficiency of the mPCR developed in this study compared to performing sPCR for 
      each virus, suggests its potential application for routine molecular diagnostic 
      purposes.
FAU - Giammarioli, Monica
AU  - Giammarioli M
AD  - Perugia, 06126, Italy.
FAU - Pellegrini, Claudia
AU  - Pellegrini C
FAU - Casciari, Cristina
AU  - Casciari C
FAU - De Mia, Gian Mario
AU  - De Mia GM
LA  - eng
PT  - Journal Article
DEP - 20071102
PL  - Switzerland
TA  - Vet Res Commun
JT  - Veterinary research communications
JID - 8100520
RN  - 0 (DNA Primers)
RN  - EC 3.1.21.- (DNA Restriction Enzymes)
SB  - IM
MH  - African Swine Fever Virus/genetics/isolation & purification
MH  - Animals
MH  - Circovirus/genetics/isolation & purification
MH  - DNA Primers/chemistry
MH  - DNA Restriction Enzymes/metabolism
MH  - Parvovirus, Porcine/genetics/isolation & purification
MH  - Polymerase Chain Reaction/methods/*veterinary
MH  - Porcine respiratory and reproductive syndrome virus/genetics/isolation & 
      purification
MH  - Sensitivity and Specificity
MH  - Swine/virology
MH  - Swine Diseases/*diagnosis/virology
MH  - Virus Diseases/diagnosis/*veterinary
MH  - Viruses/genetics/*isolation & purification
EDAT- 2007/11/03 09:00
MHDA- 2008/04/04 09:00
CRDT- 2007/11/03 09:00
PHST- 2007/07/20 00:00 [received]
PHST- 2007/10/16 00:00 [accepted]
PHST- 2007/11/03 09:00 [pubmed]
PHST- 2008/04/04 09:00 [medline]
PHST- 2007/11/03 09:00 [entrez]
AID - 10.1007/s11259-007-9026-6 [doi]
PST - ppublish
SO  - Vet Res Commun. 2008 Mar;32(3):255-62. doi: 10.1007/s11259-007-9026-6. Epub 2007 
      Nov 2.