Improved Detection Sensitivity of an Antigen Test for SARS-CoV-2 Nucleocapsid Proteins with Thio-NAD Cycling
Improved Detection Sensitivity of an Antigen Test for SARS-CoV-2 Nucleocapsid Proteins with Thio-NAD Cycling
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DOI：
10.1248/bpb.b21-00387
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发表时间：
2021-09-01
影响因子：
2
通讯作者：
ito, Etsuro
中科院分区：
文献类型：
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作者：
Kyosei, Yuta;Namba, Mayuri;ito, Etsuro
传染病的抗原检测便宜且易于使用，但检测限（LOD）通常高于被认为是金标准的PCR检测。在本研究中，我们将夹心酶联免疫吸附试验（ELISA）与硫代烟酰胺-腺嘌呤二核苷酸（thio-NAD）循环相结合，以提高2019冠状病毒病（COVID-19）抗原检测的LOD。对于严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）的重组核衣壳蛋白，我们的ELISA与硫代NAD循环的LOD为2.95 × 10（-17）摩尔/测定。当使用UV照射的无活性SARS-CoV-2时，使用我们的ELISA和硫代NAD循环获得了对应于2.6 × 10（4）RNA拷贝/测定的最小可检测病毒体。每次试验的测定体积为100 μ L。最小可检测值小于最新的SARS-CoV-2荧光免疫分析抗原检测值，表明我们的检测系统用于COVID-19诊断的有效性。
Antigen tests for infectious diseases are inexpensive and easy-to-use, but the limit of detection (LOD) is generally higher than that of PCR tests, which are considered the gold standard. In the present study, we combined a sandwich enzyme-linked immunosorbent assay (ELISA) with thionicotinamide-adenine dinucleotide (thio-NAD) cycling to improve the LOD of antigen tests for coronavirus disease 2019 (COVID-19). For recombinant nucleocapsid proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the LOD of our ELISA with thio-NAD cycling was 2.95 x 10(-17 )moles/assay. When UV-irradiated inactive SARS-CoV-2 was used, the minimum detectable virions corresponding to 2.6 x 10(4) RNA copies/assay were obtained using our ELISA with thio-NAD cycling. The assay volume for each test was 100 mu L. The minimum detectable value was smaller than that of the latest antigen test using a fluorescent immunoassay for SARS-CoV-2, indicating the validity of our detection system for COVID-19 diagnosis.