Mahallawi Waleed H
Medical Laboratory Technology Department, College of Applied Medical Sciences, Taibah University, Almadinah Almunawwarah, KSA.
J Taibah Univ Med Sci. 2021 Feb;16(1):57-62. doi: 10.1016/j.jtumed.2020.11.011. Epub 2020 Dec 31.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), can lead to severe respiratory illness. Patients with underlying comorbidities have a high risk of contracting COVID-19. Therefore, serological assays are urgently needed to diagnose asymptomatic carriers of SARS-CoV-2, to estimate the prevalence of infection, and for disease prevention and control. This study aimed to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of anti-SARS-CoV-2 antibodies in humans.
An ELISA test was designed and established to detect antibodies against the SARS-CoV-2 spike protein in serum samples from 41 quantitative reverse transcription polymerase chain reaction (qRT-PCR) - positive hospitalised COVID-19 patients. Forty-two convalescent patients' sera served as positive controls, while 117 pre-pandemic serum samples were used as negative controls.
A comparison between different SARS-CoV-2 proteins was performed, which included the full-length spike (S) protein and the S1 and S2 subunits. The full-length S protein showed the strongest reactivity for anti-SARS-CoV-2 IgG antibodies in patients' serum samples. Additionally, since antibodies could be detected at very low concentrations, the assay was found to be sensitive.
The current assay was specific, since cross-reactions with other SARS coronaviruses and respiratory viruses such as influenza were not found. Additionally, it was highly sensitive, since the test was able to identify antibodies even at very low concentrations. Therefore, this assay has promise as a screening method at the population level and may be used for in future seroepidemiological studies.
严重急性呼吸综合征冠状病毒2(SARS-CoV-2)是2019冠状病毒病(COVID-19)的病原体,可导致严重的呼吸道疾病。患有基础合并症的患者感染COVID-19的风险很高。因此,迫切需要血清学检测来诊断SARS-CoV-2无症状携带者、估计感染率以及进行疾病预防和控制。本研究旨在开发一种酶联免疫吸附测定(ELISA)法检测人抗SARS-CoV-2抗体。
设计并建立一种ELISA试验,以检测41例经定量逆转录聚合酶链反应(qRT-PCR)确诊的住院COVID-19患者血清样本中针对SARS-CoV-2刺突蛋白的抗体。42例康复患者的血清用作阳性对照,117份疫情前的血清样本用作阴性对照。
对不同的SARS-CoV-2蛋白进行了比较,包括全长刺突(S)蛋白以及S1和S2亚基。全长S蛋白在患者血清样本中对抗SARS-CoV-2 IgG抗体显示出最强的反应性。此外,由于能在极低浓度下检测到抗体,该检测方法被发现具有敏感性。
目前的检测方法具有特异性,因为未发现与其他SARS冠状病毒以及流感等呼吸道病毒发生交叉反应。此外,它具有高度敏感性,因为该检测方法即使在极低浓度下也能识别抗体。因此,该检测方法有望作为一种人群水平的筛查方法,并可能用于未来的血清流行病学研究。