Nucleoprotein-based ELISA for detection of SARS-COV-2 IgG antibodies: Could an old assay be suitable for serodiagnosis of the new coronavirus?,Journal of Virological Methods

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Nucleoprotein-based ELISA for detection of SARS-COV-2 IgG antibodies: Could an old assay be suitable for serodiagnosis of the new coronavirus?
Journal of Virological Methods ( IF 2.2 ) Pub Date : 2021-01-13 , DOI: 10.1016/j.jviromet.2021.114064
Tania Regina Tozetto-Mendoza ₁ , Kelly Aparecida Kanunfre ₂ , Lucy Santos Vilas-Boas ₁ , Evelyn Patricia Sanchez Espinoza ₃ , Heuder Gustavo Oliveira Paião ₄ , Mussya Cisotto Rocha ₂ , Anderson Vicente de Paula ₁ , Maura Salaroli de Oliveira ₅ , Daniella Bosco Zampelli ₆ , José Mauro Vieira ₆ , Lewis Buss ₇ , Silvia Figueiredo Costa ₃ , Ester Cerdeira Sabino ₈ , Steven S Witkin ₉ , Thelma Suely Okay ₁₀ , Maria Cassia Mendes-Correa ₄

Affiliation

LIM-52, Institute of Tropical Medicine, School of Medicine, Clinical Hospital, University of São Paulo, Brazil.
LIM-48, Clinical Hospital, School of Medicine, University of São Paulo, Brazil; Laboratory of Seroepidemiology and Immunobiology, Institute of Tropical Medicine, School of Medicine, University of São Paulo, Brazil.
LIM-49, Institute of Tropical Medicine, School of Medicine, University of São Paulo, Brazil; Department of Infection Diseases, Clinical Hospital, School of Medicine, University of São Paulo, Brazil.
LIM-52, Institute of Tropical Medicine, School of Medicine, Clinical Hospital, University of São Paulo, Brazil; Department of Infection Diseases, Clinical Hospital, School of Medicine, University of São Paulo, Brazil.
Department of Infection control- Clinical Hospital, School of Medicine, University of São Paulo, Brazil; Hospital Sírio Libanês, Brazil.
Hospital Sírio Libanês, Brazil.
Department of Infection Diseases, Clinical Hospital, School of Medicine, University of São Paulo, Brazil.
Department of Infection Diseases, Clinical Hospital, School of Medicine, University of São Paulo, Brazil; LIM-46, Institute of Tropical Medicine, School of Medicine, University of São Paulo, Brazil.
LIM-52, Institute of Tropical Medicine, School of Medicine, Clinical Hospital, University of São Paulo, Brazil; Department of Obstetrics and Gynecology, Weill Cornell Medicine, New York, New York, United States.
Laboratory of Seroepidemiology and Immunobiology, Institute of Tropical Medicine, School of Medicine, University of São Paulo, Brazil; Department of Pediatrics, School of Medicine, University of São Paulo, Brazil.

Objectives

We evaluated the performance of a nucleoprotein-based enzyme-linked immunosorbent assay (ELISA) for detection of IgG antibodies to SARS-CoV-2.

Methods

The ELISA was based on serum IgG reactivity to a 46-kDa protein derived from the recombinant SARS-CoV2 nucleoprotein. Assay sensitivity was assessed using serum samples from 134 COVID-19 confirmed cases obtained > 15 days after symptom onset. Specificity was determined by testing sera from 94 healthy controls. Cross-reactivity was evaluated with sera from 96 individuals with previous dengue or zika virus-confirmed infections, with 44 sera from individuals with confirmed infections to other respiratory viruses or with bacterial and fungal infections that cause pneumonia and with 40 sera negative for SARS-CoV-2 nucleoprotein by commercial ELISA kits.

Results

The majority of subjects were male and ≥ 60 years old. Assay sensitivity was 90.3 % (95 % confidence interval 84.1 %–94.2 %) and specificity was 97.9 % (92.6 %–99.4 %). There was no cross-reactivity with sera from individuals diagnosed with dengue, zika virus, influenza virus, rhinovirus, adenovirus, respiratory syncytial virus, seasonal coronavirus, Mycobacterium tuberculosis, Staphylococcus (S. aureus and coagulase-negative), Streptococcus pneumoniae, Klebsiella pneumoniae and the fungus Aspergillus fumigatus. The level of concordance of our test with results from commercial ELISA kits was 100 %.

Conclusion

The nucleoprotein-based ELISA was specific for detection of IgG anti-nucleoprotein antibodies to SARS-CoV-2. It utilizes a frequently employed low expense assay protocol and is easier to perform than other currently available commercial SARS-CoV2 antibody detection tests.

中文翻译：

基于核蛋白的 ELISA 检测 SARS-COV-2 IgG 抗体：旧的检测方法是否适用于新型冠状病毒的血清学诊断？

目标

我们评估了基于核蛋白的酶联免疫吸附测定 (ELISA) 检测 SARS-CoV-2 IgG 抗体的性能。

方法

ELISA 基于血清 IgG 对源自重组 SARS-CoV2 核蛋白的 46-kDa 蛋白的反应性。使用症状发作后 > 15 天获得的 134 例 COVID-19 确诊病例的血清样本评估了检测灵敏度。通过测试来自 94 名健康对照的血清来确定特异性。对 96 名先前已确诊登革热或寨卡病毒感染的个体的血清、44 名已确诊感染其他呼吸道病毒或患有引起肺炎的细菌和真菌感染的个体的血清以及 40 名 SARS-CoV 阴性的血清进行了交叉反应评估-2 核蛋白通过商业 ELISA 试剂盒。

结果

大多数受试者为男性且年龄≥ 60 岁。检测灵敏度为 90.3%（95% 置信区间 84.1%–94.2%），特异性为 97.9%（92.6%–99.4%）。与诊断为登革热、寨卡病毒、流感病毒、鼻病毒、腺病毒、呼吸道合胞病毒、季节性冠状病毒、结核分枝杆菌、葡萄球菌（金黄色葡萄球菌和凝固酶阴性）、肺炎链球菌、肺炎克雷伯菌的血清无交叉反应和真菌烟曲霉。我们的测试与商业 ELISA 试剂盒结果的一致性水平为 100%。