title: CFP10 and ESAT6 aptamers as effective Mycobacterial antigen diagnostic reagents.
abstract: The development of effective Mycobacterial antigen diagnostic reagents remains a high priority. The 6-kDa early secreted antigenic target (ESAT6) and 10-kDa culture filtrate protein (CFP10) are secreted early by virulent Mycobacterium tuberculosis (M. tb) and are not present in the non-virulent Bacillus Calmette-Guerin (BCG). In this study, we used a Systematic Evolution of Ligands by Exponential Enrichment (SELEX) technique to screen for a functional ssDNA aptamer "antibody" that specifically bound to ESAT6-CFP10 (CE) protein. The selected single ssDNA aptamers (CE24 and CE15) demonstrated the highest specificity and binding affinity to CFP10 (CE24: Kd = 3.75 × 10(-7) M) and ESAT6 (CE15: Kd = 1.6 × 10(-7) M). We further detected CFP10 and ESAT6 proteins in serum samples from active pulmonary tuberculosis (TB) patients, extrapulmonary TB patients and healthy donors by using an enzyme-linked oligonucleotide assay (ELONA). The results showed that the sensitivity and specificity were 100% and 94.1% (using CE24 aptamer-based ELONA) and 89.6% and 94.1% (using CE15 aptamer-based ELONA), respectively. A good correlation was observed between aptamer-based ELONA and T-SPOT TB assay. Thus, our study suggests that CE24 and CE15 have potentially broad applications as early antigen diagnostic agents not only for active pulmonary TB, extrapulmonary TB, but also possibly for latent TB infection and TB with immune-deficiency.
authors: State Key Laboratory of Virology, Department of Immunology, Hubei Province Key Laboratory of Allergy and Immunology, Wuhan University School of Basic Medical Sciences, Donghu Road 165#, Wuhan 430071, Hubei Province, China.
authors: Xiao-Lei
authors: XL
authors: Tang
authors: State Key Laboratory of Virology, Department of Immunology, Hubei Province Key Laboratory of Allergy and Immunology, Wuhan University School of Basic Medical Sciences, Donghu Road 165#, Wuhan 430071, Hubei Province, China.
authors: Ya-Xiong
authors: YX
authors: Zhou
authors: State Key Laboratory of Virology, Department of Immunology, Hubei Province Key Laboratory of Allergy and Immunology, Wuhan University School of Basic Medical Sciences, Donghu Road 165#, Wuhan 430071, Hubei Province, China; Department of Laboratory Medicine, Wuhan Medical Treatment Center, Wuhan, China.
authors: Si-Min
authors: SM
authors: Wu
authors: State Key Laboratory of Virology, Department of Immunology, Hubei Province Key Laboratory of Allergy and Immunology, Wuhan University School of Basic Medical Sciences, Donghu Road 165#, Wuhan 430071, Hubei Province, China.
authors: Qin
authors: Q
authors: Pan
authors: Department of Gastroenterology and Research of Digestive Diseases, Zhongnan Hospital, Wuhan University School of Medicine, Wuhan 430071, China.
authors: Bing
authors: B
authors: Xia
authors: State Key Laboratory of Virology, Department of Immunology, Hubei Province Key Laboratory of Allergy and Immunology, Wuhan University School of Basic Medical Sciences, Donghu Road 165#, Wuhan 430071, Hubei Province, China. Electronic address: zhangxiaolian@whu.edu.cn.
authors: Xiao-Lian
authors: XL
authors: Zhang
mesh_terms: D000328:Adult; D000942:Antigens, Bacterial; D000995:Antitubercular Agents / Q000627:therapeutic use; D052157:Aptamers, Nucleotide; D001426:Bacterial Proteins; D005260:Female; D064368:Healthy Volunteers; D006801:Humans; D055985:Latent Tuberculosis / Q000175:diagnosis* / Q000276:immunology; D008297:Male; D009169:Mycobacterium tuberculosis / Q000276:immunology*; D017711:Nonlinear Dynamics; D020411:Oligonucleotide Array Sequence Analysis; D014397:Tuberculosis, Pulmonary / Q000175:diagnosis* / Q000276:immunology
chemical_list: D000942:Antigens, Bacterial; D000995:Antitubercular Agents; D052157:Aptamers, Nucleotide; D001426:Bacterial Proteins; C117940:CFP-10 protein, Mycobacterium tuberculosis; C093276:ESAT-6 protein, Mycobacterium tuberculosis
keywords: Antigen diagnostic reagents; Aptamers; CFP10-ESAT6 (CE); Enzyme-linked oligonucleotide assay (ELONA); Tuberculosis