Introduction: A further 15 countries in Africa are predicted to be at risk of Ebola virus outbreaks in the future, as a result of climate change and vegetation. A rapid surveillance strategy is required to prevent the virus spread. A PCR alternative, Recombinase polymerase amplification (RPA), has been identified as a "game changer" for molecular diagnosis. We developed Ebola molecular assay that could detect sequence variations of the virus in 30 minutes and the test is compatible to low resource laboratories. Method: Sensitivity of our RPA-Lateral Flow was achieved by testing selected primers and probe with tenfold serial dilutions (1.34 x 1010 to 101 ) of cloned NP gene from Mayinga strain of Ebola Zaire virus in pCAGGS vector, as established by Real-Time TaqMan™ PCR. Also, tested reverse transcribed RNA from cultured Ebola Zaire virus (six different strains) - Mayinga, Gueckedou C05, Gueckedou C07, Makona, Kissidougou and Kiwit. Specificity of our assay was performed by testing Marburg, Ebola Reston and Ebola Sudan. Results: An analytical sensitivity of 1.34 x 102 was achieved, compared to a sensitivity of 1.34 x 102 for Real-Time PCR. Testing indicated the RPA-Lateral Flow could successfully detect all the Ebola Zaire virus strains, and our assay did not cross react with related viruses. Conclusion: RPA-Lateral Flow assay could detect variations in Ebola virus sequences rapidly, including the current strain that was responsible for so many deaths. Our assay sensitivity was 134 copies and it is comparable with Real-time PCR and highly specific. The assay can be performed without infrastructural complexity commonly associated with PCR.
XIX International Congress for Tropical Medicine and Malaria (ICTMM), Brisbane, Australia 18-22 September 2016
XIX International Congress for Tropical Medicine and Malaria Abstract Book / pp.797