Reverse transcription recombinase polymerase amplification assay for rapid detection of canine associated rabies virus in Africa

Rabies is a neglected disease mostly affecting the developing world. Accurate and reliable diagnostic and surveillance data forms the foundation for the formulation and monitoring of control strategies. Although various sensitive and specific tests are available for detection of rabies virus, implementation of these tests in low-resource settings are challenging and remains limited. In this study, we describe the developed of a reverse transcription recombinase polymerase amplification assay for the detection of rabies virus. The analytical sensitivity of this assay was determined to be 562 RNA copies and was performed in 20 minutes. The diagnostic sensitivity of the RT-RPA was 100% for detection of rabies virus in field samples. In conclusion, the RT-RPA assay allowed for very quick and sensitive detection of rabies virus and could be adapted for use in low-source settings.Supporting information: S1 Table. Details of rabies virus sequences used for primer and probe design for the RT-RPA assay.S2 Table. Evaluation of the mismatches between the RT-RPA primer and probe set binding regions to rabies-related lyssaviruses.The South African Research Chair initiative of the Department of Science and Technology and National Research Foundation of South Africa and the Poliomyelitis Research Foundation.http://www.plosone.orgpm2020BiochemistryGeneticsMedical VirologyMicrobiology and Plant Patholog