内部重组RBD蛋白检测SARS-CoV-2抗体免疫分析法的建立与优化

IF 0.8 Q3 MULTIDISCIPLINARY SCIENCES Makara Journal of Science Pub Date : 2022-09-30 DOI:10.7454/mss.v26i3.1342
S. P. P. Ratu, S. Mariya, R. Noviana, U. Saepuloh, H. Darusman
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引用次数: 1

摘要

由SARS-CoV-2引起的COVID-19对国际社会构成重大威胁,特别是在印度尼西亚。还实施了预防这种疾病传播的对策,包括实施一项疫苗接种规划。间接酶联免疫吸附试验(ELISA)是一种可用于分析疫苗接种后可能产生的抗体的免疫测定技术。我们制备了用于本研究的重组刺突蛋白。优化后的蛋白浓度分别为5和10 ng/mL,阻断剂浓度分别为2.5%和5%,结合物浓度分别为1:1000和1:5000。本研究的最佳条件为加标浓度为10 ng/mL,阻断剂浓度为5%,样品稀释度为1:33,偶联物浓度为1:1000。优化后的间接ELISA法检测内值为7.3,检测间值为5.3。t检验以MyBioSource市售试剂盒和免疫诊断试剂盒为参照,p值分别为0和0.313,表明本研究重组蛋白内源性ELISA试剂盒检测SARS-CoV-2抗体的能力与市售免疫诊断试剂盒相同,可用于疫苗接种后疗效评价。©2022,印度尼西亚大学。版权所有。
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Development and Optimization of an Immunoassay for the Detection of Antibodies Against SARS-CoV-2 with In-house Recombinant RBD Protein
COVID-19 caused by SARS-CoV-2 poses a major threat to the global community, particularly in Indonesia. Countermeasures to prevent the spread of this disease have also been implemented, including the implementation of a vaccination program. An immunoassay technique that can be used to analyze antibodies that might develop following vaccination is the indirect enzyme-linked immunosorbent assay (ELISA). We produced the recombinant spike protein used in this study. The optimization comprised adjusted concentrations of spike recombinant protein (5 and 10 ng/mL), blocking agent (2.5% and 5%), and conjugate (1:1000 and 1:5000). The optimal conditions in this study included a spiked concentration of 10 ng/mL, a blocking agent concentration of 5%, sample dilution of 1:33, and a conjugate concentration of 1:1000. The intra-assay value of this optimized indirect ELISA was 7.3, and the inter-assay value was 5.3. The commercial MyBioSource kit and immunodiagnostic were utilized as a reference in the T-test, with P-values of 0 and 0.313, indicating that the recombinant protein in-house ELISA kit in this study demonstrated the same ability as the commercial immunodiagnostic kit in detecting SARS-CoV-2 antibodies, allowing it to be used for post-vaccination efficacy evaluation. © 2022, Universitas Indonesia. All rights reserved.
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来源期刊
Makara Journal of Science
Makara Journal of Science MULTIDISCIPLINARY SCIENCES-
CiteScore
1.30
自引率
20.00%
发文量
24
审稿时长
24 weeks
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