PMID- 12791860
OWN - NLM
STAT- MEDLINE
DCOM- 20031001
LR  - 20240410
IS  - 0095-1137 (Print)
IS  - 1098-660X (Electronic)
IS  - 0095-1137 (Linking)
VI  - 41
IP  - 6
DP  - 2003 Jun
TI  - Development of a heteroduplex mobility assay to identify field isolates of 
      porcine reproductive and respiratory syndrome virus with nucleotide sequences 
      closely related to those of modified live-attenuated vaccines.
PG  - 2433-9
AB  - Porcine reproductive and respiratory syndrome has been devastating the swine 
      industry since the late 1980s. The disease has been controlled, to some extent, 
      through the use of modified live-attenuated (MLV) vaccines once available. 
      However, such a practice periodically resulted in isolation or detection of 
      vaccine-like viruses from pigs as determined by a partial genomic sequencing. In 
      this study, we developed a heteroduplex mobility assay (HMA) for quickly 
      identifying porcine reproductive and respiratory syndrome virus (PRRSV) isolates 
      with significant nucleotide sequence identities (>/=98%) with the modified 
      live-attenuated vaccines. The major envelope gene (ORF5) of 51 PRRSV field 
      isolates recovered before and after the introduction of the vaccines was 
      amplified, denatured, and reannealed with the HMA reference vaccine strains 
      Ingelvac PRRS MLV and Ingelvac PRRS ATP, respectively. Nine of the 51 field 
      isolates and the VR2332 parent virus of Ingelvac PRRS MLV, which were all highly 
      related to Ingelvac PRRS MLV with </=2% nucleotide sequence divergence as 
      determined by sequence analysis, were all identified by the HMA to form 
      homoduplexes with the reference Ingelvac PRRS MLV. No homoduplex-forming field 
      isolate was identified when Ingelvac PRRS ATP was used as the HMA reference 
      except for its parent virus JA142. Other field isolates with more than 2% 
      nucleotide sequence divergence with the respective reference vaccine strain 
      resulted in the formation of heteroduplexes with reduced mobility in 
      polyacrylamide gel electrophoresis. The HMA results also correlated well with the 
      results of phylogenetic analyses. The data indicated that the HMA developed in 
      the study may be a rapid and efficient method for large-scale screening of 
      potential vaccine-like PRRSV field isolates for further genetic characterization.
FAU - Key, K F
AU  - Key KF
AD  - Center for Molecular Medicine and Infectious Diseases, Department of Biomedical 
      Sciences and Pathobiology, College of Veterinary Medicine, Virginia Polytechnic 
      Institute and State University, Blacksburg, Virginia 24060-0342, USA.
FAU - Guenette, D K
AU  - Guenette DK
FAU - Yoon, K-J
AU  - Yoon KJ
FAU - Halbur, P G
AU  - Halbur PG
FAU - Toth, T E
AU  - Toth TE
FAU - Meng, X J
AU  - Meng XJ
LA  - eng
SI  - GENBANK/AF537919
SI  - GENBANK/AF537920
SI  - GENBANK/AF537921
SI  - GENBANK/AF537922
SI  - GENBANK/AF537923
SI  - GENBANK/AF537924
SI  - GENBANK/AF537925
SI  - GENBANK/AF537926
SI  - GENBANK/AF537927
SI  - GENBANK/AF537928
SI  - GENBANK/AF537929
SI  - GENBANK/AF537930
SI  - GENBANK/AF537931
SI  - GENBANK/AF537932
SI  - GENBANK/AF537933
SI  - GENBANK/AF537934
SI  - GENBANK/AF537935
SI  - GENBANK/AF537936
SI  - GENBANK/AF537937
SI  - GENBANK/AF537938
SI  - GENBANK/AF537939
SI  - GENBANK/AF537940
SI  - GENBANK/AF537941
SI  - GENBANK/AF537942
SI  - GENBANK/AF537943
SI  - GENBANK/AF537944
SI  - GENBANK/AF537945
SI  - GENBANK/AF537946
SI  - GENBANK/AF537947
SI  - GENBANK/AF537948
SI  - GENBANK/AF537949
SI  - GENBANK/AF537950
SI  - GENBANK/AF537951
SI  - GENBANK/AF537952
SI  - GENBANK/AF537953
SI  - GENBANK/AF537954
SI  - GENBANK/AF537955
SI  - GENBANK/AF537956
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Clin Microbiol
JT  - Journal of clinical microbiology
JID - 7505564
RN  - 0 (Vaccines, Attenuated)
RN  - 0 (Viral Vaccines)
SB  - IM
MH  - Animals
MH  - Heteroduplex Analysis/*methods
MH  - Molecular Sequence Data
MH  - Phylogeny
MH  - Porcine Reproductive and Respiratory Syndrome/*virology
MH  - Porcine respiratory and reproductive syndrome 
      virus/classification/genetics/*isolation & purification
MH  - Sequence Analysis, DNA
MH  - Swine
MH  - Swine Diseases/virology
MH  - *Vaccines, Attenuated
MH  - *Viral Vaccines
PMC - PMC156521
EDAT- 2003/06/07 05:00
MHDA- 2003/10/02 05:00
PMCR- 2003/06/01
CRDT- 2003/06/07 05:00
PHST- 2003/06/07 05:00 [pubmed]
PHST- 2003/10/02 05:00 [medline]
PHST- 2003/06/07 05:00 [entrez]
PHST- 2003/06/01 00:00 [pmc-release]
AID - 1457 [pii]
AID - 10.1128/JCM.41.6.2433-2439.2003 [doi]
PST - ppublish
SO  - J Clin Microbiol. 2003 Jun;41(6):2433-9. doi: 10.1128/JCM.41.6.2433-2439.2003.