Development of a Pcr and Hemi-nested Pcr Assay for the Detection of African Lyssaviruses

The genus Lyssavirus is divided into 7 genotypes. Genotype 1 is found worldwide and genotypes 2-4 are found exclusively on the African continent. Representatives of genotypes 1-4 were selected to include every lineage, group and biotype within these genotypes to account for the huge diversity observed in this genus. A reverse transcription PCR as well as a hemi-nested PCR method has been developed for the detection of all African Lyssaviruses. The nucleoprotein gene was used as the target for primers as it is well conserved and has been extensively used for genotyping isolates. This method could be used for improvement of surveillance in African countries with equipped laboratories and trained personnel when the fluorescent antibody test is not possible. The use of saliva samples of ante mortem human diagnosis as well as for samples with low viral load. Subsequent application of automated sequencing of the hemi-nested PCR product will give definitive characterization of the isolate within hours which would aid in epidemiological studies.