Aptasensor for the Detection of <i>Mycobacterium tuberculosis</i> in Sputum Utilising CFP10-ESAT6 Protein as a Selective Biomarker
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 <i>Mycobacterium tuberculosis</i> (<i>M. tuberculosis</i>) detection in clini...
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| Format: | Article |
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MDPI AG
2021-09-01
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| Series: | Nanomaterials |
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| Online Access: | https://www.mdpi.com/2079-4991/11/9/2446 |
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| author | Umi Zulaikha Mohd Azmi Nor Azah Yusof Jaafar Abdullah Faruq Mohammad Shahrul Ainliah Alang Ahmad Siti Suraiya Nurul Hanun Ahmad Raston Fatin Nabilah Mohd Faudzi Sachin K. Khiste Hamad A. Al-Lohedan |
| author_facet | Umi Zulaikha Mohd Azmi Nor Azah Yusof Jaafar Abdullah Faruq Mohammad Shahrul Ainliah Alang Ahmad Siti Suraiya Nurul Hanun Ahmad Raston Fatin Nabilah Mohd Faudzi Sachin K. Khiste Hamad A. Al-Lohedan |
| author_sort | Umi Zulaikha Mohd Azmi |
| collection | DOAJ |
| description | 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 <i>Mycobacterium tuberculosis</i> (<i>M. tuberculosis</i>) 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 Fe<sub>3</sub>O<sub>4</sub>/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 <i>M. tuberculosis</i> analysis shows good reproducibility with an relative standard deviation (RSD) of 2.5%. Further analysis of <i>M. tuberculosis</i> in sputum samples have shown good correlation with the culture method with 100% specificity and sensitivity, thus making the aptasensor a promising candidate for <i>M. tuberculosis</i> detection considering its high specificity and sensitivity with clinical samples. |
| first_indexed | 2024-03-10T07:22:24Z |
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| institution | Directory Open Access Journal |
| issn | 2079-4991 |
| language | English |
| last_indexed | 2024-03-10T07:22:24Z |
| publishDate | 2021-09-01 |
| publisher | MDPI AG |
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| series | Nanomaterials |
| spelling | doaj.art-a53d8ababb09462a96b9b32e1b5adda82023-11-22T14:32:42ZengMDPI AGNanomaterials2079-49912021-09-01119244610.3390/nano11092446Aptasensor for the Detection of <i>Mycobacterium tuberculosis</i> in Sputum Utilising CFP10-ESAT6 Protein as a Selective BiomarkerUmi Zulaikha Mohd Azmi0Nor Azah Yusof1Jaafar Abdullah2Faruq Mohammad3Shahrul Ainliah Alang Ahmad4Siti Suraiya5Nurul Hanun Ahmad Raston6Fatin Nabilah Mohd Faudzi7Sachin K. Khiste8Hamad A. Al-Lohedan9Institute of Advanced Technology, Universiti Putra Malaysia, Serdang 43400, Selangor, MalaysiaInstitute of Advanced Technology, Universiti Putra Malaysia, Serdang 43400, Selangor, MalaysiaInstitute of Advanced Technology, Universiti Putra Malaysia, Serdang 43400, Selangor, MalaysiaDepartment of Chemistry, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi ArabiaInstitute of Advanced Technology, Universiti Putra Malaysia, Serdang 43400, Selangor, MalaysiaSchool of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan 16150, MalaysiaSchool of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, UKM Bangi 43600, Selangor, MalaysiaInstitute of Advanced Technology, Universiti Putra Malaysia, Serdang 43400, Selangor, MalaysiaDepartment of Medicine, Harvard Medical School, Boston, MA 02115, USADepartment of Chemistry, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi ArabiaA 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 <i>Mycobacterium tuberculosis</i> (<i>M. tuberculosis</i>) 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 Fe<sub>3</sub>O<sub>4</sub>/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 <i>M. tuberculosis</i> analysis shows good reproducibility with an relative standard deviation (RSD) of 2.5%. Further analysis of <i>M. tuberculosis</i> in sputum samples have shown good correlation with the culture method with 100% specificity and sensitivity, thus making the aptasensor a promising candidate for <i>M. tuberculosis</i> detection considering its high specificity and sensitivity with clinical samples.https://www.mdpi.com/2079-4991/11/9/2446portable detection systemselectrochemical aptasensor<i>Mycobacterium tuberculosis</i>CFP10-ESAT6differential pulse voltammetry (DPV)iron/gold magnetic nanoparticles |
| spellingShingle | Umi Zulaikha Mohd Azmi Nor Azah Yusof Jaafar Abdullah Faruq Mohammad Shahrul Ainliah Alang Ahmad Siti Suraiya Nurul Hanun Ahmad Raston Fatin Nabilah Mohd Faudzi Sachin K. Khiste Hamad A. Al-Lohedan Aptasensor for the Detection of <i>Mycobacterium tuberculosis</i> in Sputum Utilising CFP10-ESAT6 Protein as a Selective Biomarker Nanomaterials portable detection systems electrochemical aptasensor <i>Mycobacterium tuberculosis</i> CFP10-ESAT6 differential pulse voltammetry (DPV) iron/gold magnetic nanoparticles |
| title | Aptasensor for the Detection of <i>Mycobacterium tuberculosis</i> in Sputum Utilising CFP10-ESAT6 Protein as a Selective Biomarker |
| title_full | Aptasensor for the Detection of <i>Mycobacterium tuberculosis</i> in Sputum Utilising CFP10-ESAT6 Protein as a Selective Biomarker |
| title_fullStr | Aptasensor for the Detection of <i>Mycobacterium tuberculosis</i> in Sputum Utilising CFP10-ESAT6 Protein as a Selective Biomarker |
| title_full_unstemmed | Aptasensor for the Detection of <i>Mycobacterium tuberculosis</i> in Sputum Utilising CFP10-ESAT6 Protein as a Selective Biomarker |
| title_short | Aptasensor for the Detection of <i>Mycobacterium tuberculosis</i> in Sputum Utilising CFP10-ESAT6 Protein as a Selective Biomarker |
| title_sort | aptasensor for the detection of i mycobacterium tuberculosis i in sputum utilising cfp10 esat6 protein as a selective biomarker |
| topic | portable detection systems electrochemical aptasensor <i>Mycobacterium tuberculosis</i> CFP10-ESAT6 differential pulse voltammetry (DPV) iron/gold magnetic nanoparticles |
| url | https://www.mdpi.com/2079-4991/11/9/2446 |
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