PMID- 19200443
OWN - NLM
STAT- MEDLINE
DCOM- 20090630
LR  - 20211020
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Print)
IS  - 0166-0934 (Linking)
VI  - 158
IP  - 1-2
DP  - 2009 Jun
TI  - Development of a one-step real-time quantitative PCR assay based on primer-probe 
      energy transfer for the detection of porcine reproductive and respiratory 
      syndrome virus.
PG  - 41-5
LID - 10.1016/j.jviromet.2009.01.014 [doi]
AB  - A one-step real-time RT-PCR method has been developed for the simultaneous 
      detection of both genotypes of porcine reproductive and respiratory syndrome 
      virus (PRRSV). The assay is based on primer-probe energy transfer, and the most 
      important advantage of this is the relative tolerance towards mutations in the 
      target-probe region. The primers and the probe were designed using an alignment 
      of 235 Type 1 (including all subtypes) and Type 2 PRRSV strains. According to the 
      alignment, multiple degenerations were included in the forward and reverse 
      primers to enable the detection of all PRRSV strains deposited in the GenBank. 
      Specificity was tested using 37 different PRRSV strains and eight other swine 
      pathogen viruses. The detection limit was approximately 10 copies of RNA prepared 
      from the Lelystad virus, a European Subtype 3 virus (Belarus strain Soz-8), and 
      an American vaccine virus (Ingelvac MLV). One TCID(50) was the detection limit in 
      the case of the cell cultured Lelystad virus and an American wild type isolate, 
      respectively. The melting point analysis revealed melting point decrease, but no 
      significant sensitivity and signal loss in the presence of numerous (up to five) 
      target-probe mismatches, indicating the capability of tolerating even more 
      mutations. The method was suitable for the detection and quantitation of 
      phylogenetically divergent strains and can serve as a robust, high throughput 
      tool for molecular diagnosis of the PRRSV.
FAU - Balka, Gyula
AU  - Balka G
AD  - Department of Pathology and Forensic Veterinary Medicine, Faculty of Veterinary 
      Science, Szent Istvan University, Budapest, Hungary. balka.gyula@aotk.szie.hu
FAU - Hornyak, Akos
AU  - Hornyak A
FAU - Balint, Adam
AU  - Balint A
FAU - Benyeda, Zsofia
AU  - Benyeda Z
FAU - Rusvai, Miklos
AU  - Rusvai M
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090204
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (DNA Primers)
RN  - 0 (Oligonucleotide Probes)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - DNA Primers/genetics
MH  - Fluorescence Resonance Energy Transfer/*methods
MH  - Molecular Sequence Data
MH  - Oligonucleotide Probes/genetics
MH  - Polymerase Chain Reaction/*methods
MH  - Porcine Reproductive and Respiratory Syndrome/*diagnosis
MH  - Porcine respiratory and reproductive syndrome virus/*isolation & purification
MH  - Sensitivity and Specificity
MH  - Swine
PMC - PMC7112897
EDAT- 2009/02/10 09:00
MHDA- 2009/07/01 09:00
PMCR- 2009/02/04
CRDT- 2009/02/10 09:00
PHST- 2008/09/09 00:00 [received]
PHST- 2008/11/28 00:00 [revised]
PHST- 2009/01/14 00:00 [accepted]
PHST- 2009/02/10 09:00 [entrez]
PHST- 2009/02/10 09:00 [pubmed]
PHST- 2009/07/01 09:00 [medline]
PHST- 2009/02/04 00:00 [pmc-release]
AID - S0166-0934(09)00015-9 [pii]
AID - 10.1016/j.jviromet.2009.01.014 [doi]
PST - ppublish
SO  - J Virol Methods. 2009 Jun;158(1-2):41-5. doi: 10.1016/j.jviromet.2009.01.014. 
      Epub 2009 Feb 4.