Journal
ACS APPLIED NANO MATERIALS
Volume 4, Issue 1, Pages 539-549Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsanm.0c02833
Keywords
Mycobacterium tuberculosis; CFP-10; Au@Pd nanozymes; detection probe; capturing probe; ELISA
Funding
- National Research Foundation of Korea (NRF) - Korean Government (MSIP) [NRF-2019R1A2C2007825, 2017R1A5A2015385]
- Korean Health Technology R&D Project, Ministry of Health & Welfare, Republic of Korea [HI16C1553]
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The study developed a colorimetric ELISA technique using nanozyme probes to detect CFP-10 antigens secreted by MTB for early tuberculosis diagnosis. The method showed high accuracy in discriminating active TB from other samples and has potential clinical applications.
The development of sensitive and point-of-care diagnostic tools for the serological profile of numerous Mycobacterium tuberculosis (MTB) rectifies the existing flaws in tuberculosis diagnosis. Herein, an ultrasensitive antigen-based tuberculosis detection technique and a bacterial growth monitoring strategy were designed using two nanozyme probe-based colorimetric enzyme-linked immunosorbent assay (ELISA). An anticulture filtrate protein-10 (CFP-10) factionalized magnetic nanobead (MNB) probe segregated the CFP-10 antigens from clinical samples, whereas the Au core and Pd shell (Au@Pd) nanozyme detection probe catalytically oxidized the commercially available chromogenic substrate 3,3',5,5'-tetramethylbenzidine (TMB) to yield a concentration-dependent color tonality as a signal indicator. The developed MNB-Au@Pd NZ sandwich ELISA monitors an early bacterial growth using CFP-10 antigens secreted from MTB into the liquid culture, where the limit of detection reached as low as 5.6 x 10(-12) g mL(-1) with a linear tonal range between 5.0 x 10(-13) and 5.0 x 10(-4) g mL(-1). Furthermore, active tuberculosis was discriminated from clinically isolated samples of human serum (40 samples) and urine (28 samples), with an accuracy of 92% (sensitivity: 93%; specificity: 91%) and 92% (sensitivity: 92%; specificity: 95%), respectively. This NZ-based colorimetric ELISA presents a robust and sensitive analytical technique for antigen-based clinical diagnosis, with potential applications in the clinical field, as well as for other epidemic diseases.
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