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Ebola is an emerging infectious disease caused by a deadly virus belonging to the family Filoviridae, genus  Ebolavirus . Based on their geographical distribution,  Ebolavirus  has been classified into total five species so far, mainly  Zaire ,  Sudan ,  Taï Forest ,  Bundibugyo  and  Reston . It is important to be able to differentiate the  Ebolavirus  species as they significantly differ in pathogenicity and more than one species can be present in an area. We have developed a one-step step-down RT-PCR detecting all five  Ebolavirus  species with high sensitivity (1 copy of  Ebolavirus  DNA, 10 copies of RNA and 320 copies of RNA spiked in 1 ml whole blood). The primers and FRET-probes we designed enabled us to differentiate five  Ebolavirus  species by distinct  T   m  ( Zaire : flat peaks between 53.0°C and 56.9°C;  Sudan : 51.6°C;  Reston : flat peaks between 47.5°C and 54.9°C;  Tai Forest : 52.8°C;  Bundibugyo : dual peaks at 48.9°C and 53.5°C), and by different amplicon sizes ( Zaire  255bp,  Sudan  211bp,  Reston  192bp,  Taï Forest  166bp,  Bundibugyo  146bp). This one-size-fit-all assay enables the rapid detection and discrimination of the five  Ebolavirus  species in a single reaction.

One-Step Reverse-Transcription FRET-PCR for Differential Detection of Five Ebolavirus Species