Aptasensor for the Detection of Mycobacterium tuberculosis in Sputum Utilising CFP10-ESAT6 Protein as a Selective Biomarker
Open Access
- 20 September 2021
- journal article
- research article
- Published by MDPI AG in Nanomaterials
- Vol. 11 (9), 2446
- https://doi.org/10.3390/nano11092446
Abstract
A portable electrochemical aptamer-antibody based sandwich biosensor has been designed and successfully developed using an aptamer bioreceptor immobilized onto a screen-printed electrode surface for Mycobacterium tuberculosis (M. tuberculosis) detection in clinical sputum samples. In the sensing strategy, a CFP10-ESAT6 binding aptamer was immobilized onto a graphene/polyaniline (GP/PANI)-modified gold working electrode by covalent binding via glutaraldehyde linkage. Upon interaction with the CFP10-ESAT6 antigen target, the aptamer will capture the target where the nano-labelled Fe3O4/Au MNPs conjugated antibody is used to complete the sandwich format and enhance the signal produced from the aptamer–antigen interaction. Using this strategy, the detection of CFP10-ESAT6 antigen was conducted in the concentration range of 5 to 500 ng/mL. From the analysis, the detection limit was found to be 1.5 ng/mL, thereby demonstrating the efficiency of the aptamer as a bioreceptor. The specificity study was carried out using bovine serum albumin (BSA), MPT64, and human serum, and the result demonstrated good specificity that is 7% higher than the antibody–antigen interaction reported in a previous study. The fabricated aptasensor for M. tuberculosis analysis shows good reproducibility with an relative standard deviation (RSD) of 2.5%. Further analysis of M. tuberculosis in sputum samples have shown good correlation with the culture method with 100% specificity and sensitivity, thus making the aptasensor a promising candidate for M. tuberculosis detection considering its high specificity and sensitivity with clinical samples.Funding Information
- Malaysia Research University Network (UPM/800-4/11/MRUN/2018/5539230, 9528600)
- King Saud University (Researchers Supporting Project (RSP-2021/54))
This publication has 48 references indexed in Scilit:
- Selection and Application of ssDNA Aptamers to Detect Active TB from Sputum SamplesPLOS ONE, 2012
- Rapid detection of Mycobacterium tuberculosis biomarkers in a sandwich immunoassay format using a waveguide-based optical biosensorTuberculosis, 2012
- Biosensing Technologies forMycobacterium tuberculosisDetection: Status and New DevelopmentsJournal of Immunology Research, 2011
- Dumbbell-like Au-Fe3O4 nanoparticles as label for the preparation of electrochemical immunosensorsBiosensors and Bioelectronics, 2010
- Graphene Based Electrochemical Sensors and Biosensors: A ReviewElectroanalysis, 2010
- Gold nanoparticle as an electrochemical label for inherently crosstalk-free multiplexed immunoassay on a disposable chipAnalytica Chimica Acta, 2010
- The selection and application of ssDNA aptamers against MPT64 protein in Mycobacterium tuberculosiscclm, 2009
- A General Synthetic Strategy for Oxide-Supported Metal Nanoparticle CatalystsJournal of the American Chemical Society, 2006
- In vitro selection of RNA molecules that bind specific ligandsNature, 1990
- Systematic Evolution of Ligands by Exponential Enrichment: RNA Ligands to Bacteriophage T4 DNA PolymeraseScience, 1990