Journal
CHEMOSENSORS
Volume 9, Issue 7, Pages -Publisher
MDPI
DOI: 10.3390/chemosensors9070159
Keywords
periodontal bacteria; immunoassays; quartz crystal microbalance biosensors
Funding
- OVERSEAS RESEARCH EXPERIENCE SCHOLARSHIP for Graduate Student, Graduate School and Faculty of Engineering, Chulalongkorn University
- JAPAN SCIENCE AND TECHNOLOGY AGENCY (JST) through COI [JPMJCE1305]
- THE MINISTRY OF EDUCATION, CULTURE, SPORTS, SCIENCE AND TECHNOLOGY (MEXT)
- NIPPON SHEET GLASS FOUNDATION FOR MATERIALS SCIENCE AND ENGINEERING
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In this study, a quartz crystal microbalance (QCM) immunoassay was demonstrated for quantitative assessment of periodontal bacteria, A. actinomycetemcomitans. The immunosensor constructed using a self-assembled monolayer showed a lower detection limit of 800 cells and a dynamic range up to 2.32 x 10(6) cells. This label-free method provides a sensitive approach for quantitative analysis of periodontal bacteria.
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 Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans, 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, A. actinomycetemcomitans. 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 A. actinomycetemcomitans 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 x 10(6) 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.
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