The evaluation of 7 commercial real-time PCR kits for Zaire ebolavirus using virus-like particle–encapsulated EBOV RNA

• Several real-time PCR kits for Zaire ebolavirus have been evaluated in China.
• Virus-like particles that encapsulated EBOV RNA have been prepared as positive substitutes.
• The limit of detection and the precision were completed for the assays.
• Requirements for clinical EBOV detection are basically satisfied except 1 reagent.
• It is feasible for virus-like particles as a mimic of positive samples in assay evaluation.

Several Chinese commercial real-time PCR kits for Zaire ebolavirus have been developed after the Ebola outbreak and used by Chinese medical teams in West Africa. In order to know the essential performance indicators of these kits, analytical sensitivity and precision were evaluated with virus-like particle (VLP)–encapsulated EBOV RNA. The limit of detection (LOD) and the precision were completed with a series of VLPs. The maximum and minimum of LOD was acquired by ZJ BioTech and Daan gene, respectively. For precision, all of the detection results were <5% except the maximum 5.17%. Among them, Puruikang, Daan gene, Sansure, ZJ BioTech, and Huada demonstrated superior reproducibility. Overall, the requirements of LOD <1000 copies/mL and coefficient of variation <5% could be satisfied by all kits except Kehua. Meanwhile, it is feasible for VLPs as a substitute of positive samples in assay evaluation.