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基于量子点的免疫层析试验条快速检测布氏杆菌病。

Rapid detection of brucellosis using a quantum dot-based immunochromatographic test strip.

机构信息

State Key Laboratory for Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.

Center for Disease Control and Prevention, Western Theater Command, Lanzhou, China.

出版信息

PLoS Negl Trop Dis. 2020 Sep 25;14(9):e0008557. doi: 10.1371/journal.pntd.0008557. eCollection 2020 Sep.

Abstract

Novel diagnostic tools are a major challenge for brucellosis research, especially in developing countries. Herein, we established a handheld quantum dot (QD) immunochromatographic device for the fast detection of brucellosis antibodies in the field. Total bacterial protein extracted from Brucella 104M served as labelling and coating antigen. QD labelling and immunochromatography methods were used to optimise reaction conditions, labelling conditions, reaction temperature and storage temperature. QD test strips were employed to test brucellosis serum to determine their sensitivity, specificity and stability. Test strips were compared with Rose Bengal test, standard agglutination test and colloidal gold immunochromatographic assay. Labelled Brucella total protein displayed good specificity and no cross-reactivity. The concentration of labelled total bacterial protein was 3.9 mg/ml, the coating concentration was 2.0 mg/ ml, and the serum titre with the lowest detection sensitivity was 1:25. The optimal reaction temperature for the test strip was 25-30°C. The test strip was stable after storage at room temperature and the repeatability was high, with a coefficient of variation of 4.0%. After testing 199 serum samples, the sensitivity of the QD test strip was 98.53%, the specificity was 93.57%, and the coincidence rate with the standard agglutination test was 96.98%. The developed QD immunochromatographic method can be used for rapid detection and preliminary screening of brucellosis in the field.

摘要

新型诊断工具是布鲁氏菌病研究的主要挑战,尤其是在发展中国家。在此,我们建立了一种手持式量子点(QD)免疫层析设备,用于快速检测现场的布鲁氏菌病抗体。从布鲁氏菌 104M 中提取的总细菌蛋白用作标记和包被抗原。QD 标记和免疫层析方法用于优化反应条件、标记条件、反应温度和储存温度。QD 测试条用于测试布鲁氏菌血清,以确定其灵敏度、特异性和稳定性。测试条与孟加拉玫瑰红试验、标准凝集试验和胶体金免疫层析测定法进行了比较。标记的布鲁氏菌总蛋白显示出良好的特异性和无交叉反应性。标记总细菌蛋白的浓度为 3.9mg/ml,包被浓度为 2.0mg/ml,最低检测灵敏度的血清滴度为 1:25。测试条的最佳反应温度为 25-30°C。测试条在室温下储存稳定,重复性高,变异系数为 4.0%。对 199 份血清样本进行测试后,QD 测试条的灵敏度为 98.53%,特异性为 93.57%,与标准凝集试验的符合率为 96.98%。所开发的 QD 免疫层析方法可用于现场快速检测和初步筛选布鲁氏菌病。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6a14/7540878/2f9f31d497c0/pntd.0008557.g001.jpg

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