Dielectrophoresis Concentration Method for Increased Sensitivity of the Loop-Mediated Isothermal Amplification Test for the Mycobacterium tuberculosis Complex

• Material Type: 博士論文
• Title: Dielectrophoresis Concentration Method for Increased Sensitivity of the Loop-Mediated Isothermal Amplification Test for the Mycobacterium tuberculosis Complex
• Author/Editor: 武田, 啓太
• Author Heading: 武田, 啓太
• Publication, Distribution, etc.: Austin Publishers
• Publication Date: 2021-03-22
• Publication Date (W3CDTF): 2021-03-22
• Alternative Title: 誘電泳動法を用いた結核菌濃縮法によるLoop-Mediated Isothermal Amplification法の検査感度上昇
• Periodical title: Journal of Bacteriology and Mycology
• No. or year of volume/issue: 7 8
• Volume: 7
• Issue: 8
• Pages: 1157-
• ISSN (Periodical Title): ISSN : 2471-0172
• Degree grantor/type: Nagasaki University (長崎大学)
• Date Granted: 2021-03-22
• Dissertation Number: 甲医歯薬第1344号
• Degree Type: 博士(医学)
• Text Language Code: eng
• Subject Heading: Mycobacterium Tuberculosis Complex; Dielectrophoresis; Concentration; Nucleic Acid Amplification Test
• Target Audience: 一般
• Note (General): Background: An effective concentration method is required to detect Mycobacterium tuberculosis (MTB) in paucibacillary specimens. Dielectrophoresis (DEP), a phenomenon in which a force is exerted on a dielectric particle subjected to a non-uniform electric field, is useful for concentrating bacterial specimens.Objectives: To investigate whether the procedure using the DEP method increases nucleic acid amplification test (NAAT) sensitivity.Methods: First, the capture rates were examined for multiple electrode settings by calculating the bacterial load before and after DEP using quantitative real-time PCR. These results were used to determine the optimal electrode setting. Second, conventional loop-mediated isothermal amplification (LAMP) was performed using 30 μL DNA purified using a PURE DNA extraction kit from 60 μL Mycobacterium bovis bacille de Calmette et Guérin culture fluid. The minimally-diluted specimen (theoretical concentration, 103 cfu/mL), which showed 10 consecutive negative results by LAMP, was subjected to DEP. One milliliter of the specimen was concentrated to 50 μL in the DEP chip, and LAMP was performed in 30 μL of the specimens without DNA purification.Results: At 73.2-84.9%, the capture rate was the highest in the setting with 100 kHz frequency. LAMP with DEP was performed using conventional LAMPnegative specimens, and eight of ten tests (80%) were positive. The sensitivity was higher than that of the conventional LAMP method (p = 0.0007).Conclusion: The DEP method has the potential to increase NAAT sensitivity for the MTB complex in paucibacillary specimens.; 長崎大学学位論文 学位記番号:博(医歯薬)甲第1344号 学位授与年月日:令和3年3月22日; Author: Takeda K, Chikamatsu K, Aono A, Igarashi Y, Morishige Y, Murase Y, Takaki A, Yamada H and Mitarai S; Citation: Journal of Bacteriology and Mycology, 7(8), id1157; 2020; identifier:Nagasaki University (長崎大学), 博士(医学) (2021-03-22)
• Source: https://nagasaki-u.repo.nii.ac.jp/?action=repository_action_common_download&item_id=26574&item_no=1&attribute_id=70&file_no=1 （fulltext）; https://nagasaki-u.repo.nii.ac.jp/?action=repository_action_common_download&item_id=26574&item_no=1&attribute_id=70&file_no=1
• Access Restrictions: インターネット公開
• Rights (production): Takeda et al. © All rights are reserved
• Related Material: http://hdl.handle.net/10069/00040613
• Data Provider (Database): 国立情報学研究所 : 学術機関リポジトリデータベース（IRDB）（機関リポジトリ）; https://irdb.nii.ac.jp
• Original Data Provider (Database): 長崎大学 : 長崎大学学術研究成果リポジトリ