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Aptasensor for the Detection of Mycobacterium tuberculosis in Sputum Utilising CFP10-ESAT6 Protein as a Selective Biomarker.

Authors
  • Azmi, Umi Zulaikha Mohd1
  • Yusof, Nor Azah1, 2
  • Abdullah, Jaafar1, 2
  • Mohammad, Faruq3
  • Ahmad, Shahrul Ainliah Alang1, 2
  • Suraiya, Siti4
  • Raston, Nurul Hanun Ahmad5
  • Faudzi, Fatin Nabilah Mohd1
  • Khiste, Sachin K6
  • Al-Lohedan, Hamad A3
  • 1 Institute of Advanced Technology, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia. , (Malaysia)
  • 2 Department of Chemistry, Faculty of Science, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia. , (Malaysia)
  • 3 Department of Chemistry, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia. , (Saudi Arabia)
  • 4 School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan 16150, Malaysia. , (Malaysia)
  • 5 School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, UKM Bangi 43600, Selangor, Malaysia. , (Malaysia)
  • 6 Department of Medicine, Harvard Medical School, Boston, MA 02115, USA.
Type
Published Article
Journal
Nanomaterials
Publisher
MDPI AG
Publication Date
Sep 20, 2021
Volume
11
Issue
9
Identifiers
DOI: 10.3390/nano11092446
PMID: 34578762
Source
Medline
Keywords
Language
English
License
Unknown

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.

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