Validation and performance of a multiplex serology assay to quantify antibody responses following SARS-CoV-2 infection or vaccination

IF 4.6 2区医学 Q2 IMMUNOLOGY

Clinical & Translational Immunology Pub Date : 2022-04-26 DOI:10.1002/cti2.1385

Deidre Wilkins, Anastasia A Aksyuk, Alexey Ruzin, Kevin M Tuffy, Tina Green, Rebecca Greway, Brittany Fikes, Cyrille J Bonhomme, Mark T Esser, Elizabeth J Kelly

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引用次数: 9

Abstract

Objectives

Robust, quantitative serology assays are required to accurately measure antibody levels following vaccination and natural infection. We present validation of a quantitative, multiplex, SARS-CoV-2, electrochemiluminescent (ECL) serology assay; show correlation with two established SARS-CoV-2 immunoassays; and present calibration results for two SARS-CoV-2 reference standards.

Methods

Precision, dilutional linearity, ruggedness, analytical sensitivity and specificity were evaluated. Clinical sensitivity and specificity were assessed using serum from prepandemic and SARS-CoV-2 polymerase chain reaction (PCR)-positive patient samples. Assay concordance to the established Roche Elecsys® Anti-SARS-CoV-2 immunoassay and a live-virus microneutralisation (MN) assay was evaluated.

Results

Standard curves demonstrated the assay can quantify SARS-CoV-2 antibody levels over a broad range. Assay precision (10.2−15.1% variability), dilutional linearity (≤ 1.16-fold bias per 10-fold increase in dilution), ruggedness (0.89−1.18 overall fold difference), relative accuracy (107−118%) and robust selectivity (102−104%) were demonstrated. Analytical sensitivity was 7, 13 and 7 arbitrary units mL⁻¹ for SARS-CoV-2 spike (S), receptor-binding domain (RBD) and nucleocapsid (N) antigens, respectively. For all antigens, analytical specificity was > 90% and clinical specificity was 99.0%. Clinical sensitivities for S, RBD and N antigens were 100%, 98.8% and 84.9%, respectively. Comparison with the Elecsys® immunoassay showed ≥ 87.7% agreement and linear correlation (Pearson r of 0.85, P < 0.0001) relative to the MN assay. Conversion factors for the WHO International Standard and Meso Scale Discovery® Reference Standard are presented.

Conclusions

The multiplex SARS-CoV-2 ECL serology assay is suitable for efficient, reproducible measurement of antibodies to SARS-CoV-2 antigens in human sera, supporting its use in clinical trials and sero-epidemiology studies.

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用于量化SARS-CoV-2感染或疫苗接种后抗体反应的多重血清学试验的验证和性能

目的:为了准确测量疫苗接种和自然感染后的抗体水平，需要可靠的、定量的血清学分析。我们提出了一种定量、多重、SARS-CoV-2电化学发光(ECL)血清学检测的验证;与两种已建立的SARS-CoV-2免疫分析相关;并介绍了两种SARS-CoV-2参考标准的校准结果。方法精密度、稀释线性、稳健性、分析灵敏度和特异性进行评价。使用大流行前和SARS-CoV-2聚合酶链反应(PCR)阳性患者样本的血清评估临床敏感性和特异性。对建立的罗氏Elecsys®抗sars - cov -2免疫测定法和活病毒微量中和(MN)测定法的一致性进行了评估。结果标准曲线显示，该方法可以在较宽的范围内定量SARS-CoV-2抗体水平。结果表明，检测精度(10.2 - 15.1%变异)、稀释线性(每10倍稀释增加≤1.16倍偏差)、稳健性(0.89 - 1.18倍总差)、相对准确度(107 - 118%)和稳健选择性(102 - 104%)。对SARS-CoV-2刺突(S)、受体结合域(RBD)和核衣壳(N)抗原的分析灵敏度分别为7、13和7任意单位mL−1。所有抗原的分析特异性为90%，临床特异性为99.0%。S、RBD和N抗原的临床敏感性分别为100%、98.8%和84.9%。与Elecsys®免疫测定法比较，与MN测定法的一致性≥87.7%，线性相关(Pearson r为0.85,P < 0.0001)。介绍了世界卫生组织国际标准和中尺度发现®参考标准的转换因子。结论多重SARS-CoV-2 ECL血清学检测方法适用于高效、重复性高的人血清中SARS-CoV-2抗原抗体的测定，可用于临床试验和血清流行病学研究。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Clinical & Translational Immunology Medicine-Immunology and Allergy

CiteScore

12.00

自引率

1.70%

发文量

审稿时长

13 weeks

期刊介绍： Clinical & Translational Immunology is an open access, fully peer-reviewed journal devoted to publishing cutting-edge advances in biomedical research for scientists and physicians. The Journal covers fields including cancer biology, cardiovascular research, gene therapy, immunology, vaccine development and disease pathogenesis and therapy at the earliest phases of investigation.