Label-Free Electrochemical Aptasensor for the Detection of the 3-O-C<sub>12</sub>-HSL Quorum-Sensing Molecule in <i>Pseudomonas aeruginosa</i>
<i>Pseudomonas aeruginosa</i>, an opportunistic Gram-negative bacterium, is one of the main sources of infections in healthcare environments, making its detection very important. N-3-oxo-dodecanoyl L-homoserine lactone (3-O-C<sub>12</sub>-HSL) is a characteristic molecule of...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-06-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/12/7/440 |
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| author | Denisa Capatina Teodora Lupoi Bogdan Feier Adrian Blidar Oana Hosu Mihaela Tertis Diana Olah Cecilia Cristea Radu Oprean |
| author_facet | Denisa Capatina Teodora Lupoi Bogdan Feier Adrian Blidar Oana Hosu Mihaela Tertis Diana Olah Cecilia Cristea Radu Oprean |
| author_sort | Denisa Capatina |
| collection | DOAJ |
| description | <i>Pseudomonas aeruginosa</i>, an opportunistic Gram-negative bacterium, is one of the main sources of infections in healthcare environments, making its detection very important. N-3-oxo-dodecanoyl L-homoserine lactone (3-O-C<sub>12</sub>-HSL) is a characteristic molecule of quorum sensing—a form of cell-to-cell communication between bacteria—in <i>P. aeruginosa</i>. Its detection can allow the determination of the bacterial population. In this study, the development of the first electrochemical aptasensor for the detection of 3-O-C<sub>12</sub>-HSL is reported. A carbon-based screen-printed electrode modified with gold nanoparticles proved to be the best platform for the aptasensor. Each step in the fabrication of the aptasensor (i.e., gold nanoparticles’ deposition, aptamer immobilization, incubation with the analyte) was optimized and characterized using cyclic voltammetry, differential pulse voltammetry, and electrochemical impedance spectroscopy. Different redox probes in solution were evaluated, the best results being obtained in the presence of [Fe(CN)<sub>6</sub>]<sup>4−</sup>/[Fe(CN)<sub>6</sub>]<sup>3−</sup>. The binding affinity of 106.7 nM for the immobilized thiol-terminated aptamer was determined using surface plasmon resonance. The quantification of 3-O-C<sub>12</sub>-HSL was performed by using the electrochemical signal of the redox probe before and after incubation with the analyte. The aptasensor exhibited a logarithmic range from 0.5 to 30 µM, with a limit of detection of 145 ng mL<sup>−1</sup> (0.5 µM). The aptasensor was successfully applied for the analysis of real samples (e.g., spiked urine samples, spiked microbiological growth media, and microbiological cultures). |
| first_indexed | 2024-03-09T10:22:09Z |
| format | Article |
| id | doaj.art-54a9465e3f6f4d9cadf6bd2e7d698ecd |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-09T10:22:09Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-54a9465e3f6f4d9cadf6bd2e7d698ecd2023-12-01T21:56:45ZengMDPI AGBiosensors2079-63742022-06-0112744010.3390/bios12070440Label-Free Electrochemical Aptasensor for the Detection of the 3-O-C<sub>12</sub>-HSL Quorum-Sensing Molecule in <i>Pseudomonas aeruginosa</i>Denisa Capatina0Teodora Lupoi1Bogdan Feier2Adrian Blidar3Oana Hosu4Mihaela Tertis5Diana Olah6Cecilia Cristea7Radu Oprean8Department of Analytical Chemistry, Faculty of Pharmacy, “Iuliu Hatieganu” University of Medicine and Pharmacy, 4 Pasteur Street, 400349 Cluj-Napoca, RomaniaDepartment of Analytical Chemistry, Faculty of Pharmacy, “Iuliu Hatieganu” University of Medicine and Pharmacy, 4 Pasteur Street, 400349 Cluj-Napoca, RomaniaDepartment of Analytical Chemistry, Faculty of Pharmacy, “Iuliu Hatieganu” University of Medicine and Pharmacy, 4 Pasteur Street, 400349 Cluj-Napoca, RomaniaDepartment of Analytical Chemistry, Faculty of Pharmacy, “Iuliu Hatieganu” University of Medicine and Pharmacy, 4 Pasteur Street, 400349 Cluj-Napoca, RomaniaDepartment of Analytical Chemistry, Faculty of Pharmacy, “Iuliu Hatieganu” University of Medicine and Pharmacy, 4 Pasteur Street, 400349 Cluj-Napoca, RomaniaDepartment of Analytical Chemistry, Faculty of Pharmacy, “Iuliu Hatieganu” University of Medicine and Pharmacy, 4 Pasteur Street, 400349 Cluj-Napoca, RomaniaDepartment of Infectious Diseases and Preventive Medicine, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Calea Manastur 3-5, 400372 Cluj-Napoca, RomaniaDepartment of Analytical Chemistry, Faculty of Pharmacy, “Iuliu Hatieganu” University of Medicine and Pharmacy, 4 Pasteur Street, 400349 Cluj-Napoca, RomaniaDepartment of Analytical Chemistry, Faculty of Pharmacy, “Iuliu Hatieganu” University of Medicine and Pharmacy, 4 Pasteur Street, 400349 Cluj-Napoca, Romania<i>Pseudomonas aeruginosa</i>, an opportunistic Gram-negative bacterium, is one of the main sources of infections in healthcare environments, making its detection very important. N-3-oxo-dodecanoyl L-homoserine lactone (3-O-C<sub>12</sub>-HSL) is a characteristic molecule of quorum sensing—a form of cell-to-cell communication between bacteria—in <i>P. aeruginosa</i>. Its detection can allow the determination of the bacterial population. In this study, the development of the first electrochemical aptasensor for the detection of 3-O-C<sub>12</sub>-HSL is reported. A carbon-based screen-printed electrode modified with gold nanoparticles proved to be the best platform for the aptasensor. Each step in the fabrication of the aptasensor (i.e., gold nanoparticles’ deposition, aptamer immobilization, incubation with the analyte) was optimized and characterized using cyclic voltammetry, differential pulse voltammetry, and electrochemical impedance spectroscopy. Different redox probes in solution were evaluated, the best results being obtained in the presence of [Fe(CN)<sub>6</sub>]<sup>4−</sup>/[Fe(CN)<sub>6</sub>]<sup>3−</sup>. The binding affinity of 106.7 nM for the immobilized thiol-terminated aptamer was determined using surface plasmon resonance. The quantification of 3-O-C<sub>12</sub>-HSL was performed by using the electrochemical signal of the redox probe before and after incubation with the analyte. The aptasensor exhibited a logarithmic range from 0.5 to 30 µM, with a limit of detection of 145 ng mL<sup>−1</sup> (0.5 µM). The aptasensor was successfully applied for the analysis of real samples (e.g., spiked urine samples, spiked microbiological growth media, and microbiological cultures).https://www.mdpi.com/2079-6374/12/7/440quorum sensing<i>P. aeruginosa</i>aptamerelectrochemical aptasensorelectrochemical detection3-O-C<sub>12</sub>-HSL |
| spellingShingle | Denisa Capatina Teodora Lupoi Bogdan Feier Adrian Blidar Oana Hosu Mihaela Tertis Diana Olah Cecilia Cristea Radu Oprean Label-Free Electrochemical Aptasensor for the Detection of the 3-O-C<sub>12</sub>-HSL Quorum-Sensing Molecule in <i>Pseudomonas aeruginosa</i> Biosensors quorum sensing <i>P. aeruginosa</i> aptamer electrochemical aptasensor electrochemical detection 3-O-C<sub>12</sub>-HSL |
| title | Label-Free Electrochemical Aptasensor for the Detection of the 3-O-C<sub>12</sub>-HSL Quorum-Sensing Molecule in <i>Pseudomonas aeruginosa</i> |
| title_full | Label-Free Electrochemical Aptasensor for the Detection of the 3-O-C<sub>12</sub>-HSL Quorum-Sensing Molecule in <i>Pseudomonas aeruginosa</i> |
| title_fullStr | Label-Free Electrochemical Aptasensor for the Detection of the 3-O-C<sub>12</sub>-HSL Quorum-Sensing Molecule in <i>Pseudomonas aeruginosa</i> |
| title_full_unstemmed | Label-Free Electrochemical Aptasensor for the Detection of the 3-O-C<sub>12</sub>-HSL Quorum-Sensing Molecule in <i>Pseudomonas aeruginosa</i> |
| title_short | Label-Free Electrochemical Aptasensor for the Detection of the 3-O-C<sub>12</sub>-HSL Quorum-Sensing Molecule in <i>Pseudomonas aeruginosa</i> |
| title_sort | label free electrochemical aptasensor for the detection of the 3 o c sub 12 sub hsl quorum sensing molecule in i pseudomonas aeruginosa i |
| topic | quorum sensing <i>P. aeruginosa</i> aptamer electrochemical aptasensor electrochemical detection 3-O-C<sub>12</sub>-HSL |
| url | https://www.mdpi.com/2079-6374/12/7/440 |
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