Development of a Suspension Microarray for the Genotyping of African Swine Fever Virus Targeting the SNPs in the C-Terminal End of the p72 Gene Region of the Genome
Comunicación oral en 6th EPIZONE Annual Meeting "Viruses on the move"
12 de junio de 2012
Leblanc L., Cortey M., Fernandez-Pinero J., Gallardo C., Masembe C., Okurut AR., Heath L., Van Heerden J., Sanchez-Vizcaino JM., Stahl K. y Belak S.
African swine fever virus (ASFV) causes catastrophic disease in domestic pigs, with high rates of morbidity and mortality, which can reach 100%. To date, the serological diagnosis has the drawback of not being able to differentiate variants of this virus. Previous studies have identified 22 genotypes based on sequence variation in the C-terminal region of the p72 gene, which has become the standard for categorizing ASFVs. A genotyping assay has been developed using a segment of PCR-amplified genomic DNA of approximately 450 bp, which encompasses the C-terminal end of the p72 gene. Paired DNA probes which are identical except for a single nucleotide polymorphism (SNP) at the centre position, were designed to either individually or in combination differentiate between the 22 genotypes. The assay was developed using Luminex xMAP technology. Characterization of the sample was performed by comparing fluorescence of the paired SNP probes, i.e. the probe with higher
fluorescence in a pair identified the SNP that a particular sample possessed. In the final assay, a total of 52 probes were employed, 24 SNP pairs and 4 general detection. One or more samples from each of the 22 genotypes were tested. The assay was able to detect and distinguish all 22 genotypes. Thus, the novel assay provides a powerful tool for the complex and rapid diagnosis of African swine fever
 | National Veterinary Institute (SVA). |
 | Centro de Investigación en Sanidad Animal (CISA). Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA). |
 | Makerere University (MAK). |
| Agricultural Research Council-Onderstepoort Veterinary Institute (ARC-OVI). | |
 | Departamento de Sanidad Animal. Facultad de Veterinaria. Universidad Complutense (UCM). |
| Servicio de Inmunología Viral y Medicina Preventiva (SUAT). Centro de Vigilancia Sanitaria Veterinaria (VISAVET). Universidad Complutense (UCM). |
| Swedish University of Agricultural Sciences (SLU). | |
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