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一种用于检测针对天然严重急性呼吸综合征冠状病毒2(SARS-CoV-2)刺突蛋白抗体的简单、灵敏且低成本的荧光激活细胞分选(FACS)检测方法。

A simple, sensitive, and low-cost FACS assay for detecting antibodies against the native SARS-CoV-2 spike protein.

作者信息

Hambach Julia, Stähler Tobias, Eden Thomas, Wendt Dorte, Tode Natalie, Haag Friedrich, Tolosa Eva, Altfeld Marcus, Fathi Anahita, Dahlke Christine, Addo Marylyn M, Menzel Stephan, Koch-Nolte Friedrich

机构信息

Institute of Immunology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.

Department of Virus Immunology, Heinrich Pette Institute, Leibniz Institute for Experimental Virology, Hamburg, Germany.

出版信息

Immun Inflamm Dis. 2021 Sep;9(3):905-917. doi: 10.1002/iid3.446. Epub 2021 May 12.

Abstract

BACKGROUND

Hamburg is a city state of approximately 1.9 Mio inhabitants in Northern Germany. Currently, the COVID-19 epidemic that had largely subsided during last summer is resurging in Hamburg and in other parts of the world, underlining the need for additional tools to monitor SARS-CoV-2 antibody responses.

AIM

We aimed to develop and validate a simple, low-cost assay for detecting antibodies against the native coronavirus 2 spike protein (CoV-2 S) that does not require recombinant protein or virus.

METHOD

We transiently co-transfected HEK cells or CHO cells with expression vectors encoding CoV-2 S and nuclear GFP. Spike protein-specific antibodies in human serum samples bound to transfected cells were detected with fluorochrome conjugated secondary antibodies by flow cytometry orimmunofluorescence microscopy. We applied this assay to monitor antibody development in COVID-19 patients, household contacts, and hospital personnel during the ongoing epidemic in the city state of Hamburg.

RESULTS

All recovered COVID-19 patients showed high levels of CoV-2 S-specific antibodies. With one exception, all household members that did not develop symptoms also did not develop detectable antibodies. Similarly, lab personnel that worked during the epidemic and followed social distancing guidelines remained antibody-negative.

CONCLUSION

We conclude that high-titer CoV-2 S-specific antibodies are found in most recovered COVID-19 patients and in symptomatic contacts, but only rarely in asymptomatic contacts. The assay may help health care providers to monitor disease progression and antibody responses in vaccination trials, to identify health care personnel that likely are resistant to re-infection, and recovered individuals with high antibody titers that may be suitable asplasma and/or antibody donors.

摘要

背景

汉堡是德国北部一个拥有约190万居民的城市州。目前,去年夏天基本消退的新冠疫情正在汉堡及世界其他地区卷土重来,这凸显了需要更多工具来监测严重急性呼吸综合征冠状病毒2(SARS-CoV-2)抗体反应。

目的

我们旨在开发并验证一种简单、低成本的检测方法,用于检测针对天然冠状病毒2刺突蛋白(CoV-2 S)的抗体,该方法无需重组蛋白或病毒。

方法

我们用编码CoV-2 S和核绿色荧光蛋白(GFP)的表达载体瞬时共转染人胚肾(HEK)细胞或中国仓鼠卵巢(CHO)细胞。通过流式细胞术或免疫荧光显微镜,用荧光染料偶联的二抗检测人血清样本中与转染细胞结合的刺突蛋白特异性抗体。我们应用该检测方法监测汉堡市正在流行的新冠疫情期间新冠患者、家庭接触者和医院工作人员体内抗体的产生情况。

结果

所有康复的新冠患者都显示出高水平的CoV-2 S特异性抗体。除一例例外,所有未出现症状的家庭成员也未产生可检测到的抗体。同样,在疫情期间工作并遵守社交距离准则的实验室工作人员抗体检测仍为阴性。

结论

我们得出结论,大多数康复的新冠患者和有症状的接触者体内存在高滴度的CoV-2 S特异性抗体,但无症状接触者中很少见。该检测方法可能有助于医疗保健人员在疫苗试验中监测疾病进展和抗体反应,识别可能对再次感染有抵抗力的医护人员,以及抗体滴度高且可能适合作为血浆和/或抗体供体的康复个体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/94ac/8342205/be8f8168fe00/IID3-9-905-g006.jpg

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