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在新西兰全国封锁期间,合作网络助力快速建立了针对新冠病毒的血清学检测方法。

Collaborative networks enable the rapid establishment of serological assays for SARS-CoV-2 during nationwide lockdown in New Zealand.

作者信息

McGregor Reuben, Whitcombe Alana L, Sheen Campbell R, Dickson James M, Day Catherine L, Carlton Lauren H, Sharma Prachi, Lott J Shaun, Koch Barbara, Bennett Julie, Baker Michael G, Ritchie Stephen R, Fox-Lewis Shivani, Morpeth Susan C, Taylor Susan L, Roberts Sally A, Webb Rachel H, Moreland Nicole J

机构信息

Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.

Maurice Wilkins Centre, University of Auckland, Auckland, New Zealand.

出版信息

PeerJ. 2020 Sep 3;8:e9863. doi: 10.7717/peerj.9863. eCollection 2020.

Abstract

BACKGROUND

Serological assays that detect antibodies to SARS-CoV-2 are critical for determining past infection and investigating immune responses in the COVID-19 pandemic. We established ELISA-based immunoassays using locally produced antigens when New Zealand went into a nationwide lockdown and the supply chain of diagnostic reagents was a widely held domestic concern. The relationship between serum antibody binding measured by ELISA and neutralising capacity was investigated using a surrogate viral neutralisation test (sVNT).

METHODS

A pre-pandemic sera panel ( = 113), including respiratory infections with symptom overlap with COVID-19, was used to establish assay specificity. Sera from PCR‑confirmed SARS-CoV-2 patients ( = 21), and PCR-negative patients with respiratory symptoms suggestive of COVID-19 ( = 82) that presented to the two largest hospitals in Auckland during the lockdown period were included. A two-step IgG ELISA based on the receptor binding domain (RBD) and spike protein was adapted to determine seropositivity, and neutralising antibodies that block the RBD/hACE‑2 interaction were quantified by sVNT.

RESULTS

The calculated cut-off (>0.2) in the two-step ELISA maximised specificity by classifying all pre-pandemic samples as negative. Sera from all PCR-confirmed COVID-19 patients were classified as seropositive by ELISA ≥7 days after symptom onset. There was 100% concordance between the two-step ELISA and the sVNT with all 7+ day sera from PCR‑confirmed COVID-19 patients also classified as positive with respect to neutralising antibodies. Of the symptomatic PCR-negative cohort, one individual with notable travel history was classified as positive by two-step ELISA and sVNT, demonstrating the value of serology in detecting prior infection.

CONCLUSIONS

These serological assays were established and assessed at a time when human activity was severely restricted in New Zealand. This was achieved by generous sharing of reagents and technical expertise by the international scientific community, and highly collaborative efforts of scientists and clinicians across the country. The assays have immediate utility in supporting clinical diagnostics, understanding transmission in high-risk cohorts and underpinning longer‑term 'exit' strategies based on effective vaccines and therapeutics.

摘要

背景

检测新型冠状病毒2(SARS-CoV-2)抗体的血清学检测对于确定既往感染情况以及研究新冠疫情中的免疫反应至关重要。在新西兰进入全国封锁状态且诊断试剂供应链成为国内广泛关注的问题时,我们使用本地生产的抗原建立了基于酶联免疫吸附测定(ELISA)的免疫检测方法。使用替代病毒中和试验(sVNT)研究了通过ELISA测定的血清抗体结合与中和能力之间的关系。

方法

使用一组疫情前血清(n = 113),包括与新冠症状重叠的呼吸道感染患者血清,来确定检测方法的特异性。纳入了在封锁期间就诊于奥克兰两家最大医院的经聚合酶链反应(PCR)确诊的SARS-CoV-2患者(n = 21)以及有提示新冠的呼吸道症状但PCR检测阴性的患者(n = 82)的血清。基于受体结合域(RBD)和刺突蛋白的两步IgG ELISA被用于确定血清反应阳性,通过sVNT对阻断RBD/人血管紧张素转换酶2(hACE-2)相互作用的中和抗体进行定量。

结果

两步ELISA中计算出的临界值(>0.2)通过将所有疫情前样本分类为阴性,使特异性最大化。症状出现后≥7天,所有经PCR确诊的新冠患者的血清通过ELISA均被分类为血清反应阳性。两步ELISA与sVNT之间存在100%的一致性,所有经PCR确诊的新冠患者症状出现7天及以上的血清在中和抗体方面也被分类为阳性。在有症状的PCR阴性队列中,一名有显著旅行史的个体通过两步ELISA和sVNT被分类为阳性,证明了血清学在检测既往感染方面的价值。

结论

这些血清学检测方法是在新西兰人类活动受到严格限制的时期建立和评估的。这是通过国际科学界慷慨分享试剂和技术专长以及全国科学家和临床医生的高度协作努力实现的。这些检测方法在支持临床诊断、了解高危人群中的传播以及为基于有效疫苗和治疗方法的长期“解封”策略提供依据方面具有直接效用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ddf4/7474877/5c64302f4fc5/peerj-08-9863-g001.jpg

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