Institute of Antibody Engineering, School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China.
State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Viral Hepatitis Research, Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou, China.
Anal Chem. 2020 May 19;92(10):7226-7231. doi: 10.1021/acs.analchem.0c00784. Epub 2020 May 5.
The outbreak of 2019 coronavirus disease (COVID-19) has been a challenge for hospital laboratories because of the huge number of samples that must be tested for the presence of the causative pathogen, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Simple and rapid immunodiagnostic methods are urgently needed to identify positive cases. Here we report the development of a rapid and sensitive lateral flow immunoassay (LFIA) that uses lanthanide-doped polysterene nanoparticles (LNPs) to detect anti-SARV-CoV-2 IgG in human serum. A recombinant nucleocapsid phosphoprotein of SARS-CoV-2 was dispensed onto a nitrocellulose membrane to capture specific IgG. Mouse anti-human IgG antibody was labeled with self-assembled LNPs that served as a fluorescent reporter. A 100-μL aliquot of serum samples (1:1000 dilution) was used for this assay and the whole detection process took 10 min. The results of the validation experiment met the requirements for clinical diagnostic reagents. A value of 0.0666 was defined as the cutoff value by assaying 51 normal samples. We tested 7 samples that were positive by reverse-transcription (RT-)PCR and 12 that were negative but clinically suspicious for the presence of anti-SARS-CoV-2 IgG. One of the negative samples was determined to be SARS-CoV-2 IgG positive, while the results for the other samples were consistent with those obtained by RT-PCR. Thus, this assay can achieve rapid and sensitive detection of anti-SARS-CoV-2 IgG in human serum and allow positive identification in suspicious cases; it can also be useful for monitoring the progression COVID-19 and evaluating patients' response to treatment.
2019 年冠状病毒病(COVID-19)的爆发对医院实验室来说是一个挑战,因为必须对大量样本进行检测,以确定致病病原体——严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)的存在。迫切需要简单、快速的免疫诊断方法来识别阳性病例。在这里,我们报告了一种快速、灵敏的侧向流动免疫分析(LFIA)的开发,该方法使用镧系掺杂聚苯乙烯纳米粒子(LNPs)检测人血清中的抗 SARS-CoV-2 IgG。将 SARS-CoV-2 的重组核衣壳磷酸蛋白分配到硝酸纤维素膜上以捕获特异性 IgG。小鼠抗人 IgG 抗体用自组装的 LNPs 标记,作为荧光报告物。该检测方法使用 100μL 血清样本(1:1000 稀释),整个检测过程耗时 10 分钟。验证实验的结果符合临床诊断试剂的要求。通过对 51 个正常样本进行检测,将 0.0666 定义为截断值。我们测试了 7 个 RT-PCR 阳性的样本和 12 个临床疑似但 RT-PCR 阴性的样本的抗 SARS-CoV-2 IgG。1 个阴性样本被确定为 SARS-CoV-2 IgG 阳性,而其他样本的结果与 RT-PCR 一致。因此,该检测方法可以快速、灵敏地检测人血清中的抗 SARS-CoV-2 IgG,并能在疑似病例中进行阳性鉴定;还可用于监测 COVID-19 的进展和评估患者对治疗的反应。