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使用β-丙内酯灭活、基于全病毒的间接 ELISA 检测印度 COVID-19 患者的抗 SARS-CoV-2 IgG 抗体反应。

Anti-SARS-CoV-2 IgG antibody response among Indian COVID-19 patients using β-propiolactone-inactivated, whole virus-based indirect ELISA.

机构信息

Department of Communicable Diseases, Interactive Research School for Health Affairs (IRSHA), Bharati Vidyapeeth (Deemed to be University), Pune, India.

Department of Pediatrics, Bharati Vidyapeeth Medical College, Pune, India.

出版信息

J Virol Methods. 2021 Jan;287:113996. doi: 10.1016/j.jviromet.2020.113996. Epub 2020 Oct 22.

Abstract

BACKGROUND

Coronavirus disease 2019 (COVID-19) pandemic caused by infection with severe acute respiratory syndrome - coronavirus-2 (SARS-CoV-2) continues to affect many countries and large populations. Serologic assays for antibody detection aid patient diagnosis and seroepidemiologic investigations.

METHODS

An indirect IgG ELISA was developed indigenously using β-propiolactone (BPL) inactivated SARS-CoV-2. This assay was used for screening 200 healthy donor sera collected prior to COVID-19 emergence (2017-2019), 185 serum/plasma samples of confirmed COVID-19 patients (n = 137) and 57 samples of viral RNA positive asymptomatic contacts (n = 51). The IgG response was studied in relation to duration and severity of illness.

RESULTS

The ELISA demonstrated 97 % specificity and IgG detection in >50 %, 80 %, 93.8 % and 100 % of the patients respectively during the first, second, third and fourth week of illness. IgG detection rate was higher in patients with severe disease (SD, 90.9 %) than those with mild disease (MD, 68.8 %) during the second week of illness (P = 0.027). IgG seropositivity among asymptomatic contacts was 64.7 %. IgG ELISA absorbance values were higher in SD than MD patients during the first 2 weeks of illness (P < 0.05). No significant difference was observed between the absorbance values of asymptomatic subjects and MD patients (P = 0.94).

CONCLUSION

The BPL inactivated virus-based ELISA could detect IgG antibodies early and in a significant proportion of COVID-19 patients suggesting its potential utility as a supplement to the currently used viral RNA detection tests in patient diagnosis and contact screening algorithms.

摘要

背景

由严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)感染引起的 2019 年冠状病毒病(COVID-19)大流行继续影响许多国家和大量人群。用于抗体检测的血清学检测有助于患者诊断和血清流行病学研究。

方法

我们使用β-丙内酯(BPL)灭活的 SARS-CoV-2 ,在本地开发了间接 IgG ELISA。该检测法用于筛查 200 例在 COVID-19 出现之前(2017-2019 年)采集的健康供体血清,185 例确诊 COVID-19 患者(n=137)的血清/血浆样本和 57 例病毒 RNA 阳性无症状接触者(n=51)的 57 例样本。研究了 IgG 反应与疾病持续时间和严重程度的关系。

结果

ELISA 显示,在疾病的第一、第二、第三和第四周,分别有>50%、80%、93.8%和 100%的患者出现 97%的特异性和 IgG 检测。在疾病的第二周,重症疾病(SD)患者(90.9%)的 IgG 检测率高于轻症疾病(MD)患者(68.8%)(P=0.027)。无症状接触者的 IgG 血清阳性率为 64.7%。在疾病的前 2 周,SD 患者的 IgG ELISA 吸光度值高于 MD 患者(P<0.05)。无症状受试者和 MD 患者的吸光度值之间没有观察到显著差异(P=0.94)。

结论

BPL 灭活病毒的 ELISA 能够早期检测到 COVID-19 患者的 IgG 抗体,且在很大比例的患者中均能检测到,这表明其作为目前用于患者诊断和接触者筛查算法的病毒 RNA 检测的补充具有潜在的应用价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2628/7581401/dee50c92e60f/gr1_lrg.jpg

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