PMID- 17944835
OWN - NLM
STAT- MEDLINE
DCOM- 20080408
LR  - 20211203
IS  - 0266-8254 (Print)
IS  - 0266-8254 (Linking)
VI  - 46
IP  - 1
DP  - 2008 Jan
TI  - Rapid detection and strain identification of porcine reproductive and respiratory 
      syndrome virus (PRRSV) by real-time RT-PCR.
PG  - 55-60
AB  - AIMS: To develop and validate assays based on real-time reverse 
      transcriptase-polymerase chain reaction (RT-PCR) for rapid detection and strain 
      identification (European and North American strains) of porcine reproductive and 
      respiratory syndrome virus (PRRSV) by using SYBR Green I and TaqMan probe 
      chemistries. METHODS AND RESULTS: This study describes two alternative assays 
      based on real-time RT-PCR for rapid detection and strain identification of PRRSV 
      in comparison with conventional RT-PCR. The first assay utilized SYBR Green I 
      with melting curve analysis; another assay was performed using strain-specific 
      TaqMan probes. Primers were selected from the conserved regions within ORF7 (N) 
      of both strains whereas two TaqMan probes labelled with different fluorescent 
      dyes were specifically designed for each strain. The result of strain 
      identification was confirmed by direct sequencing. Both assays can be used for 
      rapid detection and strain identification of PRRSV with a sensitivity of 10(4) 
      and 10(3) copies microl(-1) for SYBR Green and TaqMan probe, respectively. 
      CONCLUSIONS: Real-time RT-PCR is a powerful method combining rapidity, 
      specificity and efficiency for large-scale screening and strain identification of 
      PRRSV. SIGNIFICANCE AND IMPACT OF THE STUDY: The data indicate that the methods 
      developed are invaluable for detecting low levels of PRRSV infection in swine.
FAU - Lurchachaiwong, W
AU  - Lurchachaiwong W
AD  - Center of Excellence in Clinical Virology, Faculty of Medicine, Chulalongkorn 
      University, Bangkok, Thailand.
FAU - Payungporn, S
AU  - Payungporn S
FAU - Srisatidnarakul, U
AU  - Srisatidnarakul U
FAU - Mungkundar, C
AU  - Mungkundar C
FAU - Theamboonlers, A
AU  - Theamboonlers A
FAU - Poovorawan, Y
AU  - Poovorawan Y
LA  - eng
PT  - Comparative Study
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20071015
PL  - England
TA  - Lett Appl Microbiol
JT  - Letters in applied microbiology
JID - 8510094
RN  - 0 (Benzothiazoles)
RN  - 0 (DNA Primers)
RN  - 0 (Diamines)
RN  - 0 (Organic Chemicals)
RN  - 0 (Quinolines)
RN  - 0 (RNA, Viral)
RN  - 163795-75-3 (SYBR Green I)
SB  - IM
MH  - Animals
MH  - *Bacterial Typing Techniques
MH  - Benzothiazoles
MH  - DNA Primers
MH  - Diamines
MH  - Organic Chemicals
MH  - Porcine Reproductive and Respiratory Syndrome/diagnosis/*virology
MH  - Porcine respiratory and reproductive syndrome 
      virus/*classification/genetics/*isolation & purification
MH  - Quinolines
MH  - RNA, Viral/genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods
MH  - Sensitivity and Specificity
MH  - Sequence Analysis, DNA
MH  - Swine
EDAT- 2007/10/20 09:00
MHDA- 2008/04/09 09:00
CRDT- 2007/10/20 09:00
PHST- 2007/10/20 09:00 [pubmed]
PHST- 2008/04/09 09:00 [medline]
PHST- 2007/10/20 09:00 [entrez]
AID - LAM2259 [pii]
AID - 10.1111/j.1472-765X.2007.02259.x [doi]
PST - ppublish
SO  - Lett Appl Microbiol. 2008 Jan;46(1):55-60. doi: 10.1111/j.1472-765X.2007.02259.x. 
      Epub 2007 Oct 15.