PMID- 9874099
OWN - NLM
STAT- MEDLINE
DCOM- 19990208
LR  - 20220408
IS  - 0378-1135 (Print)
IS  - 1873-2542 (Electronic)
IS  - 0378-1135 (Linking)
VI  - 64
IP  - 1
DP  - 1998 Nov
TI  - Sensitive detection and typing of porcine reproductive and respiratory syndrome 
      virus by RT-PCR amplification of whole viral genes.
PG  - 7-22
AB  - Following the recent use of a live vaccine against porcine reproductive and 
      respiratory syndrome virus (PRRSV) in Denmark, both American (vaccine) and 
      European-type PRRSV now coexist in Danish herds. This situation highlighted a 
      requirement for supplementary tests for precise virus-typing. As a result, we 
      developed a RT-PCR assay able to detect as well as type PRRSV. To provide maximal 
      sequence information, complete viral open reading frames (ORFs 5 and 7) were 
      targeted for amplification. The RT-PCR test was able to amplify complete PRRSV 
      ORFs from complex materials such as boar semen containing as little as 1 TCID50 
      ml(-1) of PRRSV. Typing of viruses was accomplished by any one of three 
      strategies: (i) use of type-specific PCR primers, (ii) size determination of ORF 
      7 amplicons, (iii) DNA sequencing. All three typing strategies showed complete 
      concordance with the currently used method of typing with monoclonal antibodies 
      (MAbs) when used on a panel of PRRSV field isolates covering the period 
      1992-1997. The ORF 7-based test had particularly desirable characteristics, 
      namely, highly sensitive detection of PRRSV without apparent type bias, typing of 
      the detected virus, discrimination between pure and mixed virus populations, and 
      semi-quantitative assessment of type ratios in mixed populations, all in a single 
      PCR reaction. In addition, the obtained sequence data were used to predict two 
      simple and rapid strategies (single-enzyme restriction length polymorphy analysis 
      and oligonucleotide hybridization) for confirmation of the specificity of ORF 7 
      RT-PCR reactions. As such, the RT-PCR assay provides a new, powerful diagnostic 
      tool to study the population dynamics between present and emerging PRRSV-types.
FAU - Oleksiewicz, M B
AU  - Oleksiewicz MB
AD  - Danish Veterinary Institute for Virus Research, Lindholm, Kalvehave.
FAU - Botner, A
AU  - Botner A
FAU - Madsen, K G
AU  - Madsen KG
FAU - Storgaard, T
AU  - Storgaard T
LA  - eng
PT  - Journal Article
PL  - Netherlands
TA  - Vet Microbiol
JT  - Veterinary microbiology
JID - 7705469
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (DNA Primers)
RN  - 0 (RNA, Viral)
SB  - IM
CIN - Vet Microbiol. 2006 Jun 15;115(1-3):21-31. PMID: 16458457
MH  - Animals
MH  - Antibodies, Monoclonal
MH  - DNA Primers/chemistry
MH  - Denmark
MH  - Electrophoresis, Agar Gel/veterinary
MH  - European Union
MH  - Female
MH  - Macrophages, Alveolar
MH  - Male
MH  - Open Reading Frames
MH  - Porcine Reproductive and Respiratory Syndrome/diagnosis/*virology
MH  - Porcine respiratory and reproductive syndrome 
      virus/classification/genetics/*isolation & purification
MH  - RNA, Viral/blood
MH  - Reverse Transcriptase Polymerase Chain Reaction/veterinary
MH  - Semen/virology
MH  - Sensitivity and Specificity
MH  - Sequence Analysis, DNA
MH  - Specific Pathogen-Free Organisms
MH  - Swine
MH  - United States
PMC - PMC7117142
EDAT- 1999/01/05 00:00
MHDA- 1999/01/05 00:01
PMCR- 1998/12/10
CRDT- 1999/01/05 00:00
PHST- 1999/01/05 00:00 [pubmed]
PHST- 1999/01/05 00:01 [medline]
PHST- 1999/01/05 00:00 [entrez]
PHST- 1998/12/10 00:00 [pmc-release]
AID - S0378113598002545 [pii]
AID - S0378-1135(98)00254-5 [pii]
AID - 10.1016/s0378-1135(98)00254-5 [doi]
PST - ppublish
SO  - Vet Microbiol. 1998 Nov;64(1):7-22. doi: 10.1016/s0378-1135(98)00254-5.