In last five years, the Africa has faced two outbreaks of Zaire ebolavirus. These outbreaks have been the largest so far, and latest outbreak is still ongoing and affecting the Democratic Republic of the Congo.

We tested in parallel three different Zaire ebolavirus (EBOV) realtime RT-PCRs targeting the nucleoprotein gene (EBOV NP-RT-qPCRs) described by Trombley et al. (2010); Huang et al. (2012) and Weidmann et al. (2004). These assays are used regularly in diagnostic laboratories. The limit of detection (LOD), intra-assay repeatability using different matrixes, sensitivity and specificity were determined. In addition, the primers and probes were aligned with the sequences available in ongoing and past outbreaks in order to check the mismatches.

The specificity of all three EBOV NP-RT-qPCRs were excellent (100 %), and LODs were under or 10 copies per PCR reaction. Intra-assay repeatability was good in all assays, however the Ct-values were bit higher using the EDTA-blood based matrix. All of the primers and probes in EBOV NP-RT-qPCR assays have one or more mismatches in the probes and primers when the 2267 Zaire EBOV NP sequences, including strains Ituri from DRC outbreak (year 2018), was aligned. The EBOV strain of Bikoro (year 2018) circulating in DRC was 100 % match in Trombley and Weidmann assay, but had one mismatch in Huang assay.

在过去的五年中，非洲遭遇了两次扎伊尔埃博拉病毒的爆发。这些疫情是迄今为止最大的疫情，最近的疫情仍在持续，并影响到刚果民主共和国。

我们平行测试了针对Trombley等人描述的核蛋白基因（EBOV NP-RT-qPCR）的三种不同的扎伊尔埃博拉病毒（EBOV）实时RT-PCR。（2010）；黄等。（2012年）和魏德曼等。（2004）。这些测定法经常用于诊断实验室。确定了检测下限（LOD），使用不同基质的测定内重复性，灵敏度和特异性。另外，将引物和探针与正在进行的和过去的暴发中可用的序列比对，以检查错配。

所有三个EBOV NP-RT-qPCR的特异性均极佳（100％），每个PCR反应的LOD低于或少于10个拷贝。测定内重复性在所有测定中均良好，但使用基于EDTA血液的基质，Ct值较高。当2267 Zaire EBOV NP序列（包括来自DRC爆发的Ituri菌株（2018年））进行比对时，EBOV NP-RT-qPCR分析中的所有引物和探针在探针和引物中都有一个或多个错配。在Trombley和Weidmann分析中，在DRC中循环传播的Bikoro EBOV株（2018年）匹配率为100％，但在Huang分析中只有一个不匹配。