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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
immunology and microbiology
Design of primers and use of RT-PCR assays for typing European bluetongue virus isolates: Differentiation of field and vaccine strains
Journal of General Virology, Volume 88, No. 10, Year 2007
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Description
Bluetongue virus (BTV) is the causative agent of bluetongue, a disease of ruminant livestock that occurs almost worldwide between latitudes 35° S and 50° N. There are 24 serotypes of BTV (currently identified by serum neutralization assays). Since 1998, eight strains of six BTV serotypes (1, 2, 4, 8, 9 and 16) have invaded Europe. The most variable BTV protein is major outer-capsid component VP2, encoded by segment 2 (Seg-2) of the double-stranded RNA virus genome. VP2 represents the major target for neutralizing (and protective) antibodies that are generated in response to BTV infection, and is therefore the primary determinant of virus serotype. RT-PCR primers and assays targeting Seg-2 have been developed for rapid identification (within 24 h) of the six European BTV types. These assays are sensitive, specific and show perfect agreement with the results of conventional virus-neutralization methods. Previous studies have identified sequence variations in individual BTV genome segments that allow different isolates to be grouped on the basis of their geographical origins (topotypes). The assays described in this paper can detect any of the BTV isolates of the homologous serotype that were tested from different geographical origins (different Seg-2 topotypes). Primers were also identified that could be used to distinguish members of these different Seg-2 topotypes, as well as field and vaccine strains of most of the European BTV serotypes. The serotype-specific assays (and primers) showed no cross-amplification when they were evaluated with multiple isolates of the most closely related BTV types or with reference strains of the remaining 24 serotypes. Primers developed in this study will be updated periodically to maintain their relevance to current BTV distribution and epidemiology (http://www.iah.bbsrc.ac.uk/ dsRNA_virus_proteins/ReoID/rt-pcr-primers.htm). © 2007 SGM.
Authors & Co-Authors
Mertens, Peter Paul Clement
United Kingdom, Surrey
The Pirbright Institute
Maan, Narender Singh
United Kingdom, Surrey
The Pirbright Institute
Prasad, G.
India, Hisar
Ccs Haryana Agricultural University
Samuel, A. R.
United Kingdom, Surrey
The Pirbright Institute
Shaw, Andrew E.
United Kingdom, Surrey
The Pirbright Institute
Potgieter, Abraham Christiaan
South Africa, Onderstepoort
Onderstepoort Veterinary Institute
Anthony, Simon John
United Kingdom, Surrey
The Pirbright Institute
Maan, Sushila
United Kingdom, Surrey
The Pirbright Institute
Statistics
Citations: 116
Authors: 8
Affiliations: 3
Identifiers
Doi:
10.1099/vir.0.83023-0
ISSN:
00221317