Bundeswehr Institute of Microbiology, Neuherbergstraße 11, D-80937, Munich, Germany; German Centre for Infection Research (DZIF), Partner Site Munich, Germany.
Bundeswehr Institute of Microbiology, Neuherbergstraße 11, D-80937, Munich, Germany; German Centre for Infection Research (DZIF), Partner Site Munich, Germany.
J Virol Methods. 2021 Jun;292:114122. doi: 10.1016/j.jviromet.2021.114122. Epub 2021 Mar 8.
Reliable methods for the detection of SARS-CoV-2 neutralising antibodies (NAbs) are essential for the evaluation of vaccine candidates and for the selection of convalescent plasma donors. Virus neutralisation tests (NTs) are the gold standard for the detection and quantification of NAbs, but they are complex and require BSL3 facilities. In contrast, surrogate enzyme-linked immunosorbent assays (sELISA) offer the possibility of high-throughput testing under standard laboratory safety conditions. In this study, we investigated two commercial sELISA kits (GenScript, AdipoGen) designed for the detection of SARS-CoV-2 NAbs.
276 plasma samples were screened using commercial IgG-ELISA and NAbs titres were determined by micro-neutralisation test (micro-NT). In addition, all samples were tested in both sELISA. Sensitivity and specificity for both sELISA were determined in comparison to the micro-NT results.
57 % of the samples were SARS-CoV-2 NAb positive in micro-NT, while 43 % tested negative. Comparison with micro-NT results showed a sensitivity of 98.2 % and a specificity of 69.5 % for the GenScript ELISA. The AdipoGen ELISA had a sensitivity of 83.5 % and a specificity of 97.8 %. False negative results were obtained mainly on samples with low NAbs titres.
Both sELISA were able to qualitatively detect NAbs in plasma samples. Sensitivity and specificity differed between sELISA with GenScript superior in sensitivity and AdipoGen superior in specificity. Both sELISA were unable to quantify NAbs, thus neither of them can completely replace conventional NTs. However, in a two-step diagnostic algorithm, AdipoGen could potentially replace NT as a subsequent confirmatory test due to its high specificity but only in settings where no exact NAbs quantification is needed.
可靠的 SARS-CoV-2 中和抗体(NAb)检测方法对于评估疫苗候选物和选择恢复期血浆供体至关重要。病毒中和试验(NT)是检测和定量 NAb 的金标准,但它们复杂且需要 BSL3 设施。相比之下,替代酶联免疫吸附测定(sELISA)提供了在标准实验室安全条件下进行高通量检测的可能性。在这项研究中,我们研究了两种用于检测 SARS-CoV-2 NAb 的商业 sELISA 试剂盒(GenScript、AdipoGen)。
使用商业 IgG-ELISA 筛选了 276 份血浆样本,并通过微量中和试验(micro-NT)确定了 NAb 滴度。此外,所有样本均在两种 sELISA 中进行了测试。通过与 micro-NT 结果的比较,确定了两种 sELISA 的灵敏度和特异性。
在 micro-NT 中,57%的样本为 SARS-CoV-2 NAb 阳性,而 43%的样本为阴性。与 micro-NT 结果的比较显示,GenScript ELISA 的灵敏度为 98.2%,特异性为 69.5%。AdipoGen ELISA 的灵敏度为 83.5%,特异性为 97.8%。主要在 NAb 滴度较低的样本中获得假阴性结果。
两种 sELISA 均能够定性检测血浆样本中的 NAb。GenScript ELISA 的灵敏度和特异性优于 AdipoGen ELISA。两种 sELISA 均不能定量 NAb,因此都不能完全替代传统的 NT。然而,在两步诊断算法中,由于特异性高,AdipoGen 有可能替代 NT 作为后续的确认试验,但仅在不需要确切 NAb 定量的情况下。