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Characterization of a circulating PRRSV strain by means of random PCR cloning and full genome sequencing

Joris Van Doorsselaere UGent, Marc Geldhof UGent, Hans Nauwynck UGent and Peter Delputte UGent (2011) VIROLOGY JOURNAL. 8.
abstract
RS is a pig disease of major economic importance that causes respiratory and reproductive problems in pigs. Over the last years it has become clear that PRRSV heterogeneity is increasing. Consequently, this has a potential impact on diagnosis and strategies to counter this disease. The use of sequence-independent PCR techniques for the detection and characterization of PRRSV could be useful to bypass problems associated with the heterogeneity of this virus. A random PCR cloning approach was tested for the characterization of PRRSV strain 07V063 of unknown genetic background that circulated on a Belgian farm. By using this approach, 7305 bp of sequence data were obtained, distributed randomly across the genome. Using RT-PCR with strain-specific primers, the full length sequence (15014 nt) was obtained. Phylogenetic relationships using ORF5 and ORF1a (NSP2) sequences showed that 07V063 was classified in type 1 subtype 1 and that 07V063 was genetically different from prototype Lelystad Virus (LV). 07V063 showed 87-93% aa identity with LV ORFs coding for structural proteins. Most variation (compared to LV) was noticed in Nsp2 (81% identity) with a deletion of 28 aa. This deletion was different from other known deletions in this ORF. In conclusion, it is shown that this random PCR cloning approach can be used for the characterization of new PRRSV strains of unknown genetic background.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
EPITOPES, SUBTYPES, INFECTION, DEFINITION, HETEROGENEITY, IDENTIFICATION, LELYSTAD VIRUS, VACCINE EFFICACY, GENETIC DIVERSITY, RESPIRATORY SYNDROME VIRUS
journal title
VIROLOGY JOURNAL
Virol. J.
volume
8
article_number
160
pages
7 pages
Web of Science type
Article
Web of Science id
000290224100001
JCR category
VIROLOGY
JCR impact factor
2.343 (2011)
JCR rank
18/32 (2011)
JCR quartile
3 (2011)
ISSN
1743-422X
DOI
10.1186/1743-422X-8-160
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
1229707
handle
http://hdl.handle.net/1854/LU-1229707
date created
2011-05-23 11:12:44
date last changed
2011-05-30 14:38:47
@article{1229707,
  abstract     = {RS is a pig disease of major economic importance that causes respiratory and reproductive problems in pigs. Over the last years it has become clear that PRRSV heterogeneity is increasing. Consequently, this has a potential impact on diagnosis and strategies to counter this disease. The use of sequence-independent PCR techniques for the detection and characterization of PRRSV could be useful to bypass problems associated with the heterogeneity of this virus. A random PCR cloning approach was tested for the characterization of PRRSV strain 07V063 of unknown genetic background that circulated on a Belgian farm. By using this approach, 7305 bp of sequence data were obtained, distributed randomly across the genome. Using RT-PCR with strain-specific primers, the full length sequence (15014 nt) was obtained. Phylogenetic relationships using ORF5 and ORF1a (NSP2) sequences showed that 07V063 was classified in type 1 subtype 1 and that 07V063 was genetically different from prototype Lelystad Virus (LV). 07V063 showed 87-93\% aa identity with LV ORFs coding for structural proteins. Most variation (compared to LV) was noticed in Nsp2 (81\% identity) with a deletion of 28 aa. This deletion was different from other known deletions in this ORF. In conclusion, it is shown that this random PCR cloning approach can be used for the characterization of new PRRSV strains of unknown genetic background.},
  articleno    = {160},
  author       = {Van Doorsselaere, Joris and Geldhof, Marc and Nauwynck, Hans and Delputte, Peter},
  issn         = {1743-422X},
  journal      = {VIROLOGY JOURNAL},
  keyword      = {EPITOPES,SUBTYPES,INFECTION,DEFINITION,HETEROGENEITY,IDENTIFICATION,LELYSTAD VIRUS,VACCINE EFFICACY,GENETIC DIVERSITY,RESPIRATORY SYNDROME VIRUS},
  language     = {eng},
  pages        = {7},
  title        = {Characterization of a circulating PRRSV strain by means of random PCR cloning and full genome sequencing},
  url          = {http://dx.doi.org/10.1186/1743-422X-8-160},
  volume       = {8},
  year         = {2011},
}

Chicago
Van Doorsselaere, Joris, Marc Geldhof, Hans Nauwynck, and Peter Delputte. 2011. “Characterization of a Circulating PRRSV Strain by Means of Random PCR Cloning and Full Genome Sequencing.” Virology Journal 8.
APA
Van Doorsselaere, Joris, Geldhof, M., Nauwynck, H., & Delputte, P. (2011). Characterization of a circulating PRRSV strain by means of random PCR cloning and full genome sequencing. VIROLOGY JOURNAL, 8.
Vancouver
1.
Van Doorsselaere J, Geldhof M, Nauwynck H, Delputte P. Characterization of a circulating PRRSV strain by means of random PCR cloning and full genome sequencing. VIROLOGY JOURNAL. 2011;8.
MLA
Van Doorsselaere, Joris, Marc Geldhof, Hans Nauwynck, et al. “Characterization of a Circulating PRRSV Strain by Means of Random PCR Cloning and Full Genome Sequencing.” VIROLOGY JOURNAL 8 (2011): n. pag. Print.