Rapid Detection of IgM Antibodies against the SARS-CoV-2 Virus via Colloidal Gold Nanoparticle-Based Lateral-Flow Assay
Rapid Detection of IgM Antibodies against the SARS-CoV-2 Virus via Colloidal Gold Nanoparticle-Based Lateral-Flow Assay
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DOI：
10.1021/acsomega.0c01554
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发表时间：
2020-06-02
期刊：
影响因子：
4.1
通讯作者：
Jiao, Yong-Jun
中科院分区：
文献类型：
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作者：
Huang, Chao;Wen, Tian;Jiao, Yong-Jun
去年，新型冠状病毒病（COVID - 19）在武汉出现，并迅速传播到许多其他国家和地区。COVID - 19具有很强的人际传播传染性，可导致急性呼吸道疾病。无症状携带者能够感染其他健康人，这对公共卫生构成了挑战；世界卫生组织（WHO）已宣布COVID - 19为全球大流行疾病。核酸检测作为目前诊断COVID - 19的主要方法，可能会出现假阴性，并且由于无症状携带者的存在，难以应用于每一位疑似患者。同时，检测血液中针对新型冠状病毒（SARS - CoV - 2）的特异性抗体，如IgM抗体，是COVID - 19诊断的另一种选择，因为人们普遍认为IgM是急性感染期的一个重要指标。在本研究中，开发了一种基于胶体金纳米粒子的侧向流（AuNP - LF）检测方法，通过间接免疫层析法实现对新型冠状病毒IgM抗体的快速诊断和现场检测。为了制备AuNP - LF试纸条，将新型冠状病毒核蛋白（SARS - CoV - 2 NP）包被在分析膜上用于样本捕获，并将抗人IgM与金纳米粒子偶联形成检测报告分子。通过改变pH值和抗人IgM的量对AuNP - LF检测方法进行优化。通过检测COVID - 19患者和正常人的血清样本对AuNP - LF检测方法的性能进行评估。将结果与实时聚合酶链反应进行比较。AuNP - LF检测方法的灵敏度和特异性分别确定为100%和93.3%，卡帕统计（κ系数 = 0.872）显示出几乎完美的一致性。AuNP - LF检测方法在检测针对新型冠状病毒的IgM时表现出出色的选择性，不受其他病毒如发热伴血小板减少综合征病毒（SFTSV）和登革热病毒（DFV）的干扰。AuNP - LF检测方法能够在15分钟内得出结果，每次检测仅需10 - 20微升血清。总体而言，鉴于其具有特异性和稳定性好、操作简便、成本低以及耗时少等优点，AuNP - LF检测方法是基层医院和实验室诊断COVID - 19的一种可行方法，特别是在需要及时检测大量样本的紧急情况下。
Last year, the novel coronavirus disease (COVID-19) emerged in Wuhan, and it has rapidly spread to many other countries and regions. COVID-19 exhibits a strong human-to-human transmission infectivity and could cause acute respiratory diseases. Asymptomatic carriers are able to infect other healthy persons, and this poses a challenge for public health; the World Health Organization (WHO) has already announced COVID-19 as a global pandemic. Nucleic acid testing, considered as the current primary method for diagnosing COVID-19, might lead to false negatives and is difficult to be applied for every suspected patient because of the existence of asymptomatic carriers. Meanwhile, detecting specific antibodies in blood, such as the IgM antibody, against the SARS-CoV-2 virus is another choice for COVID-19 diagnosis, as it is widely accepted that IgM is an important indicator in the acute infection period. In this study, a colloidal gold nanoparticle-based lateral-flow (AuNP-LF) assay was developed to achieve rapid diagnosis and on-site detection of the IgM antibody against the SARS-CoV-2 virus through the indirect immunochromatography method. For preparing AuNP-LF strips, the SARS-CoV-2 nucleoprotein (SARS-CoV-2 NP) was coated on an analytical membrane for sample capture, and antihuman IgM was conjugated with AuNPs to form the detecting reporter. Optimization of AuNP-LF assay was carried out by altering the pH value and the amount of antihuman IgM. The performance of AuNP-LF assay was evaluated by testing serum samples of COVID-19 patients and normal humans. The results were compared with the real-time polymerase chain reaction. The sensitivity and specificity of AuNP-LF assay were determined to be 100 and 93.3%, respectively, and an almost perfect agreement was exhibited by Kappa statistics (. coefficient = 0.872). AuNP-LF assay showed outstanding selectivity in the detection of IgM against the SARS-CoV-2 virus with no interference from other viruses such as severe fever with thrombocytopenia syndrome virus (SFTSV) and dengue virus (DFV). AuNP-LF assay was able to achieve results within 15 min and needed only 10-20 mu L serum for each test. As a whole, in the light of its advantages such as excellent specificity and stability, easy operation, low cost, and being less time-consuming, AuNP-LF assay is a feasible method for the diagnosis of COVID-19 in primary hospitals and laboratories, especially in emergency situations in which numerous samples need to be tested on time.