PMID- 20713151
OWN - NLM
STAT- MEDLINE
DCOM- 20110215
LR  - 20101101
IS  - 1096-1194 (Electronic)
IS  - 0890-8508 (Linking)
VI  - 24
IP  - 6
DP  - 2010 Dec
TI  - PriProET based melting point analyses on PRRSV positive field samples.
PG  - 411-4
LID - 10.1016/j.mcp.2010.08.002 [doi]
AB  - A one-step real time RT-PCR method has previously been developed for the 
      simultaneous detection of both genotypes of porcine reproductive and respiratory 
      syndrome virus (PRRSV). For further evaluation of the assay and a detailed 
      characterization of the probe binding sites a collection of 24 PRRSV positive 
      field samples from Hungary, Serbia, Austria, a highly pathogenic strain from 
      Bhutan and commercially available MLV vaccine strains were collected and 
      sequenced from the terminal part of ORF6 to the 3' end UTR. The regions that were 
      targeted by the probe were analyzed in detail, and their sequences were compared 
      to that of the probe. Each sample showed a positive result with the PriProET 
      assay, and the samples that showed nucleotide mismatches on the probe binding 
      region had shifted melting points compared to the perfectly matching Lelystad 
      strain. Based on the melting temperatures the strains were classified into 8 
      groups ranging from 62.4 degrees C to 75.5 degrees C. The samples with the lowest melting 
      temperatures were Type I strains which had less mismatches on the probe binding 
      site than Type II strains. However, these mutations were closer to the 3' end of 
      the probe. It can be speculated that mismatches near the 5' end of the probe had 
      lower influence on the melting temperature.
CI  - Copyright (c) 2010 Elsevier Ltd. All rights reserved.
FAU - Balka, Gyula
AU  - Balka G
AD  - Department of Pathology and Forensic Veterinary Medicine, Faculty of Veterinary 
      Science, Szent Istvan University, Istvan u. 2, H-1078 Budapest, Hungary. 
      balka.gyula@aotk.szie.hu
FAU - Hornyak, Akos
AU  - Hornyak A
FAU - Dan, Adam
AU  - Dan A
FAU - Ladinig, Andrea
AU  - Ladinig A
FAU - Biksi, Imre
AU  - Biksi I
FAU - Rusvai, Miklos
AU  - Rusvai M
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20100814
PL  - England
TA  - Mol Cell Probes
JT  - Molecular and cellular probes
JID - 8709751
RN  - 0 (DNA Primers)
RN  - 0 (DNA, Viral)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - DNA Primers/*metabolism
MH  - DNA, Viral/*genetics
MH  - *Energy Transfer
MH  - Molecular Sequence Data
MH  - Nucleic Acid Denaturation
MH  - Porcine respiratory and reproductive syndrome virus/*genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods
MH  - Transition Temperature
EDAT- 2010/08/18 06:00
MHDA- 2011/02/16 06:00
CRDT- 2010/08/18 06:00
PHST- 2010/02/26 00:00 [received]
PHST- 2010/07/28 00:00 [revised]
PHST- 2010/08/04 00:00 [accepted]
PHST- 2010/08/18 06:00 [entrez]
PHST- 2010/08/18 06:00 [pubmed]
PHST- 2011/02/16 06:00 [medline]
AID - S0890-8508(10)00051-4 [pii]
AID - 10.1016/j.mcp.2010.08.002 [doi]
PST - ppublish
SO  - Mol Cell Probes. 2010 Dec;24(6):411-4. doi: 10.1016/j.mcp.2010.08.002. Epub 2010 
      Aug 14.