Quantitative Assessment of Periodontal Bacteria Using a Cell-Based Immunoassay with Functionalized Quartz Crystal Microbalance
Periodontal disease is an inflammatory disorder that is triggered by bacterial plaque and causes the destruction of the tooth-supporting tissues leading to tooth loss. Several bacteria species, including <i>Porphyromonas gingivalis</i> and <i>Aggregatibacter actinomycetemcomitans&l...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-06-01
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| Series: | Chemosensors |
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| Online Access: | https://www.mdpi.com/2227-9040/9/7/159 |
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| author | Satit Rodphukdeekul Miyuki Tabata Chindanai Ratanaporncharoen Yasuo Takeuchi Pakpum Somboon Watcharee Boonlue Yuji Miyahara Mana Sriyudthsak |
| author_facet | Satit Rodphukdeekul Miyuki Tabata Chindanai Ratanaporncharoen Yasuo Takeuchi Pakpum Somboon Watcharee Boonlue Yuji Miyahara Mana Sriyudthsak |
| author_sort | Satit Rodphukdeekul |
| collection | DOAJ |
| description | Periodontal disease is an inflammatory disorder that is triggered by bacterial plaque and causes the destruction of the tooth-supporting tissues leading to tooth loss. Several bacteria species, including <i>Porphyromonas gingivalis</i> and <i>Aggregatibacter actinomycetemcomitans</i>, are considered to be associated with severe periodontal conditions. In this study, we demonstrated a quartz crystal microbalance (QCM) immunoassay for quantitative assessment of the periodontal bacteria, <i>A</i><i>. actinomycetemcomitans</i>. An immunosensor was constructed using a self-assembled monolayer of 11-mercaptoundecanoic acid (11-MUA) on the gold surface of a QCM chip. The 11-MUA layer was evaluated using a cyclic voltammetry technique to determine its mass and packing density. Next, a monoclonal antibody was covalently linked to 11-MUA using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide to act as the biorecognition element. The specificity of the monoclonal antibody was confirmed by an enzyme-linked immunosorbent assay. A calibration curve, for the relationship between the frequency shifts and number of bacteria, was used to calculate the number of <i>A</i><i>. actinomycetemcomitans</i> bacteria in a test sample. Based on a regression equation, the lower detection limit was 800 cells, with a dynamic range up to 2.32 × 10<sup>6</sup> cells. Thus, the QCM biosensor in this study provides a sensitive and label-free method for quantitative analysis of periodontal bacteria. The method can be used in various biosensing assays for practical application and routine detection of periodontitis pathogens. |
| first_indexed | 2024-03-10T10:03:38Z |
| format | Article |
| id | doaj.art-7cbda45b322d44e195212f0c896e579f |
| institution | Directory Open Access Journal |
| issn | 2227-9040 |
| language | English |
| last_indexed | 2024-03-10T10:03:38Z |
| publishDate | 2021-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Chemosensors |
| spelling | doaj.art-7cbda45b322d44e195212f0c896e579f2023-11-22T01:43:36ZengMDPI AGChemosensors2227-90402021-06-019715910.3390/chemosensors9070159Quantitative Assessment of Periodontal Bacteria Using a Cell-Based Immunoassay with Functionalized Quartz Crystal MicrobalanceSatit Rodphukdeekul0Miyuki Tabata1Chindanai Ratanaporncharoen2Yasuo Takeuchi3Pakpum Somboon4Watcharee Boonlue5Yuji Miyahara6Mana Sriyudthsak7Biomedical Engineering Program, Faculty of Engineering, Chulalongkorn University, Bangkok 10330, ThailandInstitute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo 101-0062, JapanInstitute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo 101-0062, JapanDepartment of Periodontology, Faculty of Dentistry, Tokyo Medical and Dental University, Tokyo 113-8510, JapanBiomedical Engineering Program, Faculty of Engineering, Chulalongkorn University, Bangkok 10330, ThailandDepartment of Nutrition and Dietetics, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok 10330, ThailandInstitute of Biomaterials and Bioengineering, Tokyo Medical and Dental University, Tokyo 101-0062, JapanDepartment of Electrical Engineering, Faculty of Engineering, Chulalongkorn University, Bangkok 10330, ThailandPeriodontal disease is an inflammatory disorder that is triggered by bacterial plaque and causes the destruction of the tooth-supporting tissues leading to tooth loss. Several bacteria species, including <i>Porphyromonas gingivalis</i> and <i>Aggregatibacter actinomycetemcomitans</i>, are considered to be associated with severe periodontal conditions. In this study, we demonstrated a quartz crystal microbalance (QCM) immunoassay for quantitative assessment of the periodontal bacteria, <i>A</i><i>. actinomycetemcomitans</i>. An immunosensor was constructed using a self-assembled monolayer of 11-mercaptoundecanoic acid (11-MUA) on the gold surface of a QCM chip. The 11-MUA layer was evaluated using a cyclic voltammetry technique to determine its mass and packing density. Next, a monoclonal antibody was covalently linked to 11-MUA using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide to act as the biorecognition element. The specificity of the monoclonal antibody was confirmed by an enzyme-linked immunosorbent assay. A calibration curve, for the relationship between the frequency shifts and number of bacteria, was used to calculate the number of <i>A</i><i>. actinomycetemcomitans</i> bacteria in a test sample. Based on a regression equation, the lower detection limit was 800 cells, with a dynamic range up to 2.32 × 10<sup>6</sup> cells. Thus, the QCM biosensor in this study provides a sensitive and label-free method for quantitative analysis of periodontal bacteria. The method can be used in various biosensing assays for practical application and routine detection of periodontitis pathogens.https://www.mdpi.com/2227-9040/9/7/159periodontal bacteriaimmunoassaysquartz crystal microbalance biosensors |
| spellingShingle | Satit Rodphukdeekul Miyuki Tabata Chindanai Ratanaporncharoen Yasuo Takeuchi Pakpum Somboon Watcharee Boonlue Yuji Miyahara Mana Sriyudthsak Quantitative Assessment of Periodontal Bacteria Using a Cell-Based Immunoassay with Functionalized Quartz Crystal Microbalance Chemosensors periodontal bacteria immunoassays quartz crystal microbalance biosensors |
| title | Quantitative Assessment of Periodontal Bacteria Using a Cell-Based Immunoassay with Functionalized Quartz Crystal Microbalance |
| title_full | Quantitative Assessment of Periodontal Bacteria Using a Cell-Based Immunoassay with Functionalized Quartz Crystal Microbalance |
| title_fullStr | Quantitative Assessment of Periodontal Bacteria Using a Cell-Based Immunoassay with Functionalized Quartz Crystal Microbalance |
| title_full_unstemmed | Quantitative Assessment of Periodontal Bacteria Using a Cell-Based Immunoassay with Functionalized Quartz Crystal Microbalance |
| title_short | Quantitative Assessment of Periodontal Bacteria Using a Cell-Based Immunoassay with Functionalized Quartz Crystal Microbalance |
| title_sort | quantitative assessment of periodontal bacteria using a cell based immunoassay with functionalized quartz crystal microbalance |
| topic | periodontal bacteria immunoassays quartz crystal microbalance biosensors |
| url | https://www.mdpi.com/2227-9040/9/7/159 |
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