Examples of the HIV-EBOV RNA standard values using different technologies.

Quantitative RT-PCR (qRT-PCR) was performed in duplicate in three independent experiments using HIV-1 LTR specific primers and probe; samples were quantified against the 3rd HIV-1 International standard (assigned value 185 000 IU/mL) run in parallel. The same samples and set of primers and probe were used in a droplet digital RT-PCR (ddRT-PCR) and results expressed as average of two independent experiments run in duplicate. HIV-EBOV RNA preparations were also quantified using Ebola virus np or I specific primers and probes (qRT-PCR np/l) and copies per mL were inferred using standard curves obtained by serial dilution of the lentiviral plasmids used to produce the particles. Ebola virus np-specific primers and probe were also used in a ddRT-PCR. Both HIV-EBOV RNA high titre preparations were also analysed by NTA. Results are reported as average concentration±standard deviation calculated on 5 acquisitions of a 1:10 dilution in universal buffer. Where results are reported as ‘copies/mL’, the relationship to genuine genome equivalence numbers is unknown.Examples of the HIV-EBOV RNA standard values using different technologies.