PMID- 7814546
OWN - NLM
STAT- MEDLINE
DCOM- 19950206
LR  - 20210526
IS  - 0095-1137 (Print)
IS  - 1098-660X (Electronic)
IS  - 0095-1137 (Linking)
VI  - 32
IP  - 9
DP  - 1994 Sep
TI  - Detection of porcine reproductive and respiratory syndrome virus and efficient 
      differentiation between Canadian and European strains by reverse transcription 
      and PCR amplification.
PG  - 2197-203
AB  - Two sets of oligonucleotide primers (1008PS-1009PR and 1010PLS-1011PLR) were 
      designed according to the sequence of the nucleocapsid protein (N) gene of Quebec 
      reference strain IAF-exp91 of porcine reproductive and respiratory syndrome virus 
      (PRRSV). The primers were used in reverse transcription and PCR (RT-PCR) 
      experiments for detection of viral genomic RNA either from infected porcine 
      alveolar macrophages (PAM) or tissues from experimentally infected 
      specific-pathogen-free pigs. Considering the high degree of variation detected 
      between the nucleotide sequences of the N genes of IAF-exp91 and Lelystad virus 
      (LV) strains of PRRSV, the primers 1008PS-1009PR were referred to as the specific 
      primers, since they were chosen in such a manner that they could amplify only 
      sequences from IAF-exp91 RNA and not from LV. On the other hand, the primer pair 
      1010PLS-1011PLR was common to both strains of PRRSV. When analyzed by agarose gel 
      electrophoresis, the products of RT-PCR from each set of primers were resolved as 
      single band of the predicted size, the specificity of amplified products being 
      confirmed by Southern blotting with a specific IAF-exp91 N gene probe. No 
      amplification was observed when RNA was extracted from uninfected PAM or from 
      other porcine viruses. As expected, only the common primer pair was able to 
      amplify RNA from the Quebec reference strain and two European strains (LV and 
      Weybridge). The resulting bands displayed differences in electrophoretic 
      mobilities due to the absence of 37 nucleotides in both European strains, thus 
      allowing their differentiation from the IAF-exp91 strain. Most of the tissue 
      culture-adapted Quebec isolates were detected with both primer pairs. The 
      sensitivity of the enzymatic amplification method for detection of PRRSV from 
      lung tissues was a 50% tissue culture infective dose of 5. RT-PCR was found to be 
      more sensitive than indirect immunofluorescence assay for detection of PRRSV in 
      tissues from experimentally infected pigs and as sensitive as virus isolation in 
      PAM, especially when combined with Southern blotting with the digoxigenin-labeled 
      N probe and chemiluminescence detection.
FAU - Mardassi, H
AU  - Mardassi H
AD  - Centre de Recherche en Virologie, Institut Armand-Frappier, Universite du Quebec, 
      Laval, Canada.
FAU - Wilson, L
AU  - Wilson L
FAU - Mounir, S
AU  - Mounir S
FAU - Dea, S
AU  - Dea S
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Clin Microbiol
JT  - Journal of clinical microbiology
JID - 7505564
RN  - 0 (DNA Primers)
RN  - 0 (RNA, Viral)
SB  - IM
MH  - Abortion, Veterinary/epidemiology/*microbiology
MH  - Animals
MH  - Base Sequence
MH  - Blotting, Southern
MH  - Capsid
MH  - Cells, Cultured
MH  - DNA Primers
MH  - Europe/epidemiology
MH  - Female
MH  - Macrophages, Alveolar/*virology
MH  - Molecular Sequence Data
MH  - Open Reading Frames
MH  - *Polymerase Chain Reaction
MH  - Pregnancy
MH  - Quebec/epidemiology
MH  - RNA Viruses/*isolation & purification
MH  - RNA, Viral/*genetics
MH  - Respiratory Tract Infections/epidemiology/microbiology/*veterinary
MH  - Species Specificity
MH  - Specific Pathogen-Free Organisms
MH  - Swine
MH  - Swine Diseases/epidemiology/*microbiology
PMC - PMC263966
EDAT- 1994/09/01 00:00
MHDA- 1994/09/01 00:01
PMCR- 1994/09/01
CRDT- 1994/09/01 00:00
PHST- 1994/09/01 00:00 [pubmed]
PHST- 1994/09/01 00:01 [medline]
PHST- 1994/09/01 00:00 [entrez]
PHST- 1994/09/01 00:00 [pmc-release]
AID - 10.1128/jcm.32.9.2197-2203.1994 [doi]
PST - ppublish
SO  - J Clin Microbiol. 1994 Sep;32(9):2197-203. doi: 10.1128/jcm.32.9.2197-2203.1994.