Introducing a portable electrochemical biosensor for Mycobacterium avium subsp. paratuberculosis detection using graphene oxide and chitosan
Abstract In this contribution, a novel, low-cost, high throughput, and ultra-selective electrochemical DNA nanobiosensor was developed for accurate on-site detection of Mycobacterium avium subspecies paratuberculosis (MAP) in real media for practical diagnosis of Johne's disease (JD). The metho...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2024-01-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-50706-z |
| _version_ | 1797363369530884096 |
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| author | Nahid Naghshgar Saied Hosseinzadeh Abdollah Derakhshandeh Ruhollah Shaali Mohammad Mahdi Doroodmand |
| author_facet | Nahid Naghshgar Saied Hosseinzadeh Abdollah Derakhshandeh Ruhollah Shaali Mohammad Mahdi Doroodmand |
| author_sort | Nahid Naghshgar |
| collection | DOAJ |
| description | Abstract In this contribution, a novel, low-cost, high throughput, and ultra-selective electrochemical DNA nanobiosensor was developed for accurate on-site detection of Mycobacterium avium subspecies paratuberculosis (MAP) in real media for practical diagnosis of Johne's disease (JD). The method was designed based on the immobilization of graphene oxide and chitosan biopolymer on the surface of a glassy carbon electrode, modified by electrochemical immobilization of graphene oxide and chitosan biopolymer, followed by activation of biopolymer via 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxy succinimide (EDC/NHS) coupling system. Afterward, the commercial probe DNA (ssDNA) was stabilized on the activated electrode surface to prepare an ultra-selective ssDNA-stabilized nanobiosensor for MAP sensing called “ssDNA-stabilized GO-CH-EDC/NHS-modified electrode”. Several characterization methods distinguished the bioelectrode. The DNA hybridization between the nanobiosensor and target DNA was confirmed by cyclic voltammetry and differential pulse voltammetry. "At optimal experimental conditions, the nanobiosensor showed a linear range of 1.0 × 10−15–1.0 × 10−12 mol L−1, a detection limit as low as 1.53 × 10−13 mol L−1, and a repeatability with a relative standard deviation (%RSD) of 4.7%. The reproducibility was also appropriate, with a %RSD of about 10%. It was used to diagnose MAP in real samples with highly accurate results. Therefore, the developed nanobiosensor can be used for clinical diagnosis of MAP. |
| first_indexed | 2024-03-08T16:19:23Z |
| format | Article |
| id | doaj.art-649ffe8e41e243bd975f9b80ca58f2c2 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-08T16:19:23Z |
| publishDate | 2024-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-649ffe8e41e243bd975f9b80ca58f2c22024-01-07T12:25:35ZengNature PortfolioScientific Reports2045-23222024-01-0114111310.1038/s41598-023-50706-zIntroducing a portable electrochemical biosensor for Mycobacterium avium subsp. paratuberculosis detection using graphene oxide and chitosanNahid Naghshgar0Saied Hosseinzadeh1Abdollah Derakhshandeh2Ruhollah Shaali3Mohammad Mahdi Doroodmand4Department of Pathobiology, School of Veterinary Medicine, Shiraz UniversityDepartment of Food Hygiene and Public Health, School of Veterinary Medicine, Shiraz UniversityDepartment of Pathobiology, School of Veterinary Medicine, Shiraz UniversityDepartment of Chemistry, College of Science, Shiraz UniversityDepartment of Chemistry, College of Science, Shiraz UniversityAbstract In this contribution, a novel, low-cost, high throughput, and ultra-selective electrochemical DNA nanobiosensor was developed for accurate on-site detection of Mycobacterium avium subspecies paratuberculosis (MAP) in real media for practical diagnosis of Johne's disease (JD). The method was designed based on the immobilization of graphene oxide and chitosan biopolymer on the surface of a glassy carbon electrode, modified by electrochemical immobilization of graphene oxide and chitosan biopolymer, followed by activation of biopolymer via 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxy succinimide (EDC/NHS) coupling system. Afterward, the commercial probe DNA (ssDNA) was stabilized on the activated electrode surface to prepare an ultra-selective ssDNA-stabilized nanobiosensor for MAP sensing called “ssDNA-stabilized GO-CH-EDC/NHS-modified electrode”. Several characterization methods distinguished the bioelectrode. The DNA hybridization between the nanobiosensor and target DNA was confirmed by cyclic voltammetry and differential pulse voltammetry. "At optimal experimental conditions, the nanobiosensor showed a linear range of 1.0 × 10−15–1.0 × 10−12 mol L−1, a detection limit as low as 1.53 × 10−13 mol L−1, and a repeatability with a relative standard deviation (%RSD) of 4.7%. The reproducibility was also appropriate, with a %RSD of about 10%. It was used to diagnose MAP in real samples with highly accurate results. Therefore, the developed nanobiosensor can be used for clinical diagnosis of MAP.https://doi.org/10.1038/s41598-023-50706-z |
| spellingShingle | Nahid Naghshgar Saied Hosseinzadeh Abdollah Derakhshandeh Ruhollah Shaali Mohammad Mahdi Doroodmand Introducing a portable electrochemical biosensor for Mycobacterium avium subsp. paratuberculosis detection using graphene oxide and chitosan Scientific Reports |
| title | Introducing a portable electrochemical biosensor for Mycobacterium avium subsp. paratuberculosis detection using graphene oxide and chitosan |
| title_full | Introducing a portable electrochemical biosensor for Mycobacterium avium subsp. paratuberculosis detection using graphene oxide and chitosan |
| title_fullStr | Introducing a portable electrochemical biosensor for Mycobacterium avium subsp. paratuberculosis detection using graphene oxide and chitosan |
| title_full_unstemmed | Introducing a portable electrochemical biosensor for Mycobacterium avium subsp. paratuberculosis detection using graphene oxide and chitosan |
| title_short | Introducing a portable electrochemical biosensor for Mycobacterium avium subsp. paratuberculosis detection using graphene oxide and chitosan |
| title_sort | introducing a portable electrochemical biosensor for mycobacterium avium subsp paratuberculosis detection using graphene oxide and chitosan |
| url | https://doi.org/10.1038/s41598-023-50706-z |
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