An aptasensor for the detection of Mycobacterium tuberculosis secreted immunogenic protein MPT64 in clinical samples towards tuberculosis detection

Marzhan Sypabekova, Kanat Dukenbayev, Anna Tsepke, Akmaral Akisheva, Nurlan Oralbayev, Damira Kanayeva

Research output: Contribution to journalArticle

Abstract

This work presents experimental results on detection of Mycobacterium tuberculosis secreted protein MPT64 using an interdigitated electrode (IDE) which acts as a platform for capturing an immunogenic protein and an electrochemical impedance spectroscopy (EIS) as a detection technique. The assay involves a special receptor, single stranded DNA (ssDNA) aptamer, which specifically recognizes MPT64 protein. The ssDNA immobilization on IDE was based on a co-adsorbent immobilization at an optimized ratio of a 1/100 HS-(CH6)6-OP(O)2O-(CH2CH2O)6-5′-TTTTT-aptamer-3′/6-mercaptohexanol. The optimal sample incubation time required for a signal generation on an aptamer modified IDE was found to be at a range of 15–20 min. Atomic Force Microscopy (AFM) results confirmed a possible formation of an aptamer - MPT64 complex with a 20 nm roughness on the IDE surface vs. 4.5 nm roughness for the IDE modified with the aptamer only. A limit of detection for the EIS aptasensor based on an IDE for the detection of MPT64 in measurement buffer was 4.1 fM. The developed EIS aptasensor was evaluated on both serum and sputum clinical samples from the same TB (−) and TB (+) patients having a specificity and sensitivity for the sputum sample analysis 100% and 76.47%, respectively, and for the serum sample analysis 100% and 88.24%, respectively. The developed aptasensor presents a sensitive method for the TB diagnosis with the fast detection time.
Original languageEnglish
Article number16273
Number of pages11
JournalScientific Reports
Volume9
DOIs
Publication statusPublished - Nov 7 2019

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Mycobacterium tuberculosis
Electrodes
Tuberculosis
Dielectric Spectroscopy
Proteins
Electrochemical impedance spectroscopy
HS 6
Single-Stranded DNA
Sputum
Immobilization
DNA
Surface roughness
Nucleotide Aptamers
Atomic Force Microscopy
Serum
Adsorbents
Limit of Detection
Assays
Atomic force microscopy
Buffers

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biomedical Engineering

Cite this

An aptasensor for the detection of Mycobacterium tuberculosis secreted immunogenic protein MPT64 in clinical samples towards tuberculosis detection. / Sypabekova, Marzhan; Dukenbayev, Kanat; Tsepke, Anna; Akisheva, Akmaral ; Oralbayev, Nurlan; Kanayeva, Damira.

In: Scientific Reports, Vol. 9, 16273, 07.11.2019.

Research output: Contribution to journalArticle

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