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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
Real Time RT-PCR Assays for Detection and Typing of African Horse Sickness Virus
PLoS ONE, Volume 9, No. 4, Article e93758, Year 2014
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Description
Although African horse sickness (AHS) can cause up to 95% mortality in horses, naïve animals can be protected by vaccination against the homologous AHSV serotype. Genome segment 2 (Seg-2) encodes outer capsid protein VP2, the most variable of the AHSV proteins. VP2 is also a primary target for AHSV specific neutralising antibodies, and consequently determines the identity of the nine AHSV serotypes. In contrast VP1 (the viral polymerase) and VP3 (the sub-core shell protein), encoded by Seg-1 and Seg-3 respectively, are highly conserved, representing virus species/orbivirus-serogroup-specific antigens. We report development and evaluation of real-time RT-PCR assays targeting AHSV Seg-1 or Seg-3, that can detect any AHSV type (virus species/serogroup-specific assays), as well as type-specific assays targeting Seg-2 of the nine AHSV serotypes. These assays were evaluated using isolates of different AHSV serotypes and other closely related orbiviruses, from the 'Orbivirus Reference Collection' (ORC) at The Pirbright Institute. The assays were shown to be AHSV virus-species-specific, or type-specific (as designed) and can be used for rapid, sensitive and reliable detection and identification (typing) of AHSV RNA in infected blood, tissue samples, homogenised Culicoides, or tissue culture supernatant. None of the assays amplified cDNAs from closely related heterologous orbiviruses, or from uninfected host animals or cell cultures. © 2014 Bachanek-Bankowska et al.
Available Materials
https://efashare.b-cdn.net/share/pmc/articles/PMC3983086/bin/pone.0093758.s001.doc
Authors & Co-Authors
Bachanek-Bankowska, Katarzyna
United Kingdom, Surrey
The Pirbright Institute
Maan, Sushila
United Kingdom, Surrey
The Pirbright Institute
India, Hisar
Lala Lajpat Rai University of Veterinary and Animal Sciences
Castillo-Olivares, Javier
United Kingdom, Surrey
The Pirbright Institute
Manning, Nicola M.
United Kingdom, Surrey
The Pirbright Institute
Maan, Narender Singh
United Kingdom, Surrey
The Pirbright Institute
India, Hisar
Lala Lajpat Rai University of Veterinary and Animal Sciences
Potgieter, Abraham Christiaan
South Africa, Centurion
Deltamune Pty Ltd
South Africa, Potchefstroom
North-west University
Di Nardo, Antonello
United Kingdom, Surrey
The Pirbright Institute
Sutton, Geoff
United Kingdom, Oxford
The Wellcome Centre for Human Genetics
Batten, Carrie Anne
United Kingdom, Surrey
The Pirbright Institute
Mertens, Peter Paul Clement
United Kingdom, Surrey
The Pirbright Institute
Statistics
Citations: 30
Authors: 10
Affiliations: 5
Identifiers
Doi:
10.1371/journal.pone.0093758
e-ISSN:
19326203