serotype specific primers and gel-based rt-pcr assays for ‘typing’ african horse sickness virus identification of strains from africa血清型特异性引物和凝胶rt - pcr检测u201c打字u201d非洲马瘟病毒株来自非洲的识别.pdfVIP

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serotype specific primers and gel-based rt-pcr assays for ‘typing’ african horse sickness virus identification of strains from africa血清型特异性引物和凝胶rt - pcr检测u201c打字u201d非洲马瘟病毒株来自非洲的识别.pdf

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serotype specific primers and gel-based rt-pcr assays for ‘typing’ african horse sickness virus identification of strains from africa血清型特异性引物和凝胶rt - pcr检测u201c打字u201d非洲马瘟病毒株来自非洲的识别

Serotype Specific Primers and Gel-Based RT-PCR Assays for ‘Typing’ African Horse Sickness Virus: Identification of Strains from Africa ¤ Narender S. Maan , Sushila Maan, Kyriaki Nomikou, Manjunatha N. Belaganahalli, Katarzyna Bachanek- Bankowska, Peter P. C. Mertens* Vector-borne Disease Programme, Institute for Animal Health, Pirbright Laboratory, Pirbright, Woking, Surrey, United Kingdom Abstract African horse sickness is a devastating, transboundary animal disease, that is ‘listed’ by the Office International des Epizooties (OIE). Although attenuated, inactivated and subunit vaccines have been developed for African horse sickness virus (AHSV), these are serotype-specific and their effective deployment therefore relies on rapid and reliable identification of virus type. AHSV serotype is controlled by the specificity of interactions between neutralising antibodies, and components of the outer-capsid, particularly protein VP2 (encoded by AHSV genome segment 2 (Seg-2)). We report the development and evaluation of novel gel based reverse transcription-PCR (RT–PCR) assays targeting AHSV Seg-2, which can be used to very significantly increase the speed and reliability of detection and identification (compared to virus neutralisation tests) of the nine serotypes of AHSV. Primer sets were designed targeting regions of Seg-2 that are conserved between strains within each of the AHSV serotype (types 1 to 9). These assays were evaluated using multiple AHSV strains from the orbivirus reference collection at IAH (/dsRNA_virus_proteins/ReoID/AHSV-isolates.htm). In each case the Seg-2 primers showed a high level of specificity and failed to cross-amplify the most closely related heterologous AHSV types, or other related orbiviruses (such as bluetongue viru