Detection of Mycobacterium tuberculosis CFP10 with visual biosensor based on Aptamer and AuNCs
LI Duo1, LI Min1, GUO Jingwei2, LIU Miao3, Tan Yunhong2, HUANG Haowen4, YUAN Shishan1
1. School of Medicine, Hunan Normal University, Changsha 410013, China; 2. Laboratory Department of Hunan Chest Hospital, Changsha 410081, China; 3. Department of Laboratory Medicine, Hunan Provincial People's Hospital, Changsha 410005, China; 4. School of Chemistry and Chemical Engineering, Hunan University of Science and Technology, Xiangtan 411201, China
Abstract：Objective A visual biosensor based on aptamer of CFP10 and AuNCs mimetic enzyme was established to detect Mycobacterium tuberculosis CFP10 in serum of tuberculosis patients and provide a new strategy for assisting tuberculosis diagnosis. Methods The colloidal solution of AuNCs was prepared by glutathione reduction method and the peroxidase like catalytic activity of AuNCs was confirmed by the TMB colorimetric method; The binding ability of CFP10 aptamer modified by sulfhydryl to AuNCs was verified by using the biotin-avidin-HRP system; The CFP10 was captured by the microwell plate coated with the CFP10 polyclonal antibody and then capture aptamer-AuNCs to construct a gold nanocluster visual biosensor for detecting CPF10; The sensitivity, specificity, negative predictive value, positive predictive value and diagnostic efficiency of the visual biosensor were evaluated. Results The synthesized AuNCs exhibit peroxidase like activity and does not affect their activity when coupled with aptamers. Compared with the control group, the presence of CFP10 caused the color of GNPs to turn blue. As the concentration of CFP10 increases, this change process can be clearly observed with the naked eye. The sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic efficiency of the visual biosensor for detecting serum CFP10 in diagnosing Mycobacterium tuberculosis infection were 94.74%, 75.86%, 83.72%, 91.67%, and 86.56%, respectively. Conclusion The Visual biosensors based on Aptamer and AuNCs can detect CFP10 and assist the diagnosis of Mycobacterium tuberculosis infection.
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2024, Vol. 21 
