PMID- 9925208
OWN - NLM
STAT- MEDLINE
DCOM- 19990415
LR  - 20190522
IS  - 1040-6387 (Print)
IS  - 1040-6387 (Linking)
VI  - 11
IP  - 1
DP  - 1999 Jan
TI  - Detection of porcine reproductive and respiratory syndrome virus by reverse 
      transcription-polymerase chain reaction using different regions of the viral 
      genome.
PG  - 27-33
AB  - Serologic studies have revealed strain variability between American and European 
      isolates and among American isolates of porcine reproductive and respiratory 
      syndrome virus (PRRSV). The objective of this study was to develop an assay for 
      the routine diagnosis of PRRSV in field specimens using reverse 
      transcription-polymerase chain reaction (RT-PCR) amplification of conserved 
      genomic regions. Twenty-four field isolates of PRRSV from different regions of 
      the USA were analyzed in the study. Six primer pairs from open reading frames 
      (ORFs) 4, 6, and 7 of the American strain (ATCC VR-2332) and from ORF 1b of the 
      Lelystad strain were used for the amplification of the viral genome by PCR. 
      Amplification products of the expected sizes were obtained from all isolates by 
      PCR amplification of ORF 7, the gene encoding the nucleocapsid protein. 
      Oligonucleotide primers designed to amplify ORFs 4 and 6 detected 92% and 96% of 
      the isolates, respectively, whereas primers for the amplification of ORF 1b 
      detected 88% of all isolates. The specificity of the amplified products of ORF 7 
      from 7 field isolates and 2 reference strains was confirmed by chemiluminescent 
      hybridization using an internal digoxigenin-labeled DNA probe. Sequence analysis 
      of this region indicated variation in the nucleotide sequence of 2 isolates that 
      did not hybridize with the internal probe. These results indicate that ORF 7 may 
      serve as a potential target for the detection of PRRSV strains by RT-PCR and that 
      genomic variability should be considered when nucleic acid hybridization is used 
      to confirm the specificity of PCR amplification for diagnostic purposes.
FAU - Guarino, H
AU  - Guarino H
AD  - Department of Veterinary Diagnostic Medicine, College of Veterinary Medicine, 
      University of Minnesota, St. Paul 55108, USA.
FAU - Goyal, S M
AU  - Goyal SM
FAU - Murtaugh, M P
AU  - Murtaugh MP
FAU - Morrison, R B
AU  - Morrison RB
FAU - Kapur, V
AU  - Kapur V
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Vet Diagn Invest
JT  - Journal of veterinary diagnostic investigation : official publication of the 
      American Association of Veterinary Laboratory Diagnosticians, Inc
JID - 9011490
RN  - 0 (DNA Primers)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - DNA Primers
MH  - *Genome, Viral
MH  - Lung/*virology
MH  - Macrophages, Alveolar/*virology
MH  - Porcine Reproductive and Respiratory Syndrome/blood/*diagnosis/virology
MH  - Porcine respiratory and reproductive syndrome virus/genetics/*isolation & 
      purification
MH  - Reverse Transcriptase Polymerase Chain Reaction/methods/*veterinary
MH  - Sensitivity and Specificity
MH  - Swine
MH  - United States
MH  - Viral Load
EDAT- 1999/01/30 00:00
MHDA- 1999/01/30 00:01
CRDT- 1999/01/30 00:00
PHST- 1999/01/30 00:00 [pubmed]
PHST- 1999/01/30 00:01 [medline]
PHST- 1999/01/30 00:00 [entrez]
AID - 10.1177/104063879901100104 [doi]
PST - ppublish
SO  - J Vet Diagn Invest. 1999 Jan;11(1):27-33. doi: 10.1177/104063879901100104.