研究生: |
謝承恩 Cheng-En Hsieh |
---|---|
論文名稱: |
糖尿病與類風濕性關節炎病患之紫膜生物光電晶片檢測分析 Detection of diabetics and rheumatoid arthritis using purple membrane-based photoelectric chips |
指導教授: |
陳秀美
Hsiu-Mei Chen |
口試委員: |
林景堉
Ching-Yu Lin 張哲菖 Che-Chang Chang 吳雪霞 Hsueh-Hsia Wu 葉旻鑫 Min-Hsin Yeh |
學位類別: |
碩士 Master |
系所名稱: |
工程學院 - 化學工程系 Department of Chemical Engineering |
論文出版年: | 2019 |
畢業學年度: | 107 |
語文別: | 中文 |
論文頁數: | 124 |
中文關鍵詞: | 紫膜 、生物光電晶片 、糖尿病檢測 、類風濕性關節炎檢測 |
外文關鍵詞: | purple membrane, biochip, Diabetics detection, Rheumatoid arthritis detection |
相關次數: | 點閱:311 下載:0 |
分享至: |
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報 |
現今社會有許多文明病,其中糖尿病及類風濕性關節炎較為常見,而這兩種疾病的確認都需要至醫院進行繁瑣檢驗,所以開發快速且方便的簡易檢測法將有其優勢。本論文中,這兩種疾病的檢測晶片皆是延續先前實驗室所開發之製程,以古生嗜鹽菌中紫膜所含的細菌視紫質作為訊號轉換器,並且利用細菌視紫質的光電流響應和入射光強度成正相關的原理製作而成。糖尿病的檢測是利用可分別檢測非糖化血紅素 (HbA0) 與糖化血紅素 (HbA1c) 之Hb aptamer-PM及HbA1c aptamer-PM複合晶片對19個全血檢體 (分別來自14位糖尿病患者及5位正常人) 進行檢測,再與毛細管電泳檢測出之HbA1c %數值做比較,發現兩者間的Pearson相關係數為0.981,證明所開發晶片對實際檢體的檢測能力。對晶片的專一性及保存性進行測試,發現這兩種晶片間並無交叉檢測,且保存28天後仍保有至少94%檢測活性。此外,以可檢測自體免疫抗體的ITIH3542-556 Cit peptide-PM複合晶片進行類風濕性關節炎分析,對40個血清檢體 (分別來自20位類風濕性關節炎患者及20位正常人) 進行檢測,並接續與anti-IgA-AuNPs結合,發現與anti-IgA-AuNPs結合前後的數據與anti-CCP2標準檢測法之結果間的Pearson相關係數分別為-0.839與-0.836,證明ITIH3542-556 Cit peptide-PM複合晶片可以單一步驟直接檢測類風濕性關節炎。此外,此晶片經保存28天後,其檢測活性並無絲毫下降,說明其具有高穩定性,適合商業化應用。
Both diabetes and rheumatoid arthritis are common civilized diseases and their diagnoses generally rely on cumbersome hospital tests. Therefore, it will be beneficial to develop a convenient, fast, and simple detection method for each disease. This thesis continued using the detection chips of those two diseases previously developed in this laboratory, employing bacteriorhodopsin (BR) residing in Halobacterium salinarum purple membrane (PM) as the signal transducer as well as the positive correlation between the BR photocurrent and the illumination intensity. For diabetes detection, 19 whole blood samples collected from 14 diabetics and 5 healthy patients were each analyzed with hemoglobin (Hb) aptamer-PM and glycated hemoglobin (HbA1c) aptamer-PM chips, which specifically recognized non-glycated Hb (HbA0) and HbA1c, respectively. Compared with the HbA1c value analyzed using the capillary electrophoresis method, a Pearson correlation coefficient of 0.981 was obtained, demonstrating the application potential of those two PM-based chips in clinical detection. No cross reactivity was observed between these two chips and both chips retained at least 94% of their detection activity after 28-day storage. For rheumatoid arthritis detection, an ITIH3542-556 Cit peptide-PM chip that recognized autoimmune antibodies was used to analyze 40 serum samples collected from 20 rheumatoid arthritis and 20 healthy patients. The serum-coated chips were further labeled with anti-IgA-AuNPs. The results showed that Pearson correlation coefficients of -0.839 and -0.836 were obtained before and after the anti-IgA-AuNPs coating step, respectively, in comparison with the results acquired using the standard anti-CCP2 detection method. Therefore, a single-step detection of rheumatoid arthritis can be achieved using the ITIH3542-556 Cit peptide-PM chip. Furthermore, the detection chip remained fully active after 28-day storage, revealing its high stability and suitability for commercial applications.
American Diabetes Association. Diagnosis and classification of diabetes mellitus. Diabetes Care. 2014, S81-S90.
American Diabetes Association. Diagnosis and classification of diabetes mellitus. Diabetes Care. 2004, S5-S10.
Balashov SP. Protonation reactions and their coupling in bacteriorhodopsin. Biochimica et Biophysica Acta. 2000, 1460:75-94.
Bunn HF, Haney DN, Gabbay KH, Gallop PM. Further identification of the nature and linkage of the carbohydrate in hemoglobinA1c. Biochemical and Biophysical Research Communications. 1975, 67:103-109.
Chang KW, Li J, Yang CH, Shiesh SC, Lee GB. An integrated microfluidic system for measurement of glycated hemoglobin levels by using an aptamer-antibody assay on magnetic beads. Biosensors and Bioelectronics. 2015, 68:397-403.
Chen HM, Lin CJ, Jheng KR, Kosasih A, Chang JY. Effect of graphene oxide on affinity-immobilization of purple membranes on solid supports. Colloids Surf B Biointerfaces. 2014, 116:482-488.
English E, John WG, Milosevich ET. In vitro determination of hemoglobin A1c for diabetes diagnosis and management: technology update. Pathology and Laboratory Medicine International. 2014, 21-31.
Gonzalez O, Gronau S, Falb M, Pfeiffer F, Mendoza E, Zimmer R, Oesterhelt D. Reconstruction, modeling & analysis of Halobacterium salinarum R-1 metabolism. Molecular BioSystems. 2007, 148-159.
Greiner A, Plischke H, Kellner H, Gruber R. Association of anti-cyclic citrullinated peptide antibodies, anti-citrullin antibodies, and IgM and IgA rheumatoid factors with serological parameters of disease activity in rheumatoid arthritis. Annals of the New York Academy of Sciences. 2005, 1050:295-303.
Hirschfield GM, Pepys MB. C-reactive protein: a critical update. Journal of Clinical Investigation. 2003, 111:1805-1812.
Koivunen ME, Krogsrud RL. Principles of Immunochemical Techniques Used in Clinical Laboratories. Laboratory Medicine 2006, 490-497.
Lin CY et al., Comparative analysis of novel autoantibody isotypes against citrullinated-inter-alpha-trypsin inhibitor heavy chain 3 (ITIH3)542–556 peptide in serum from Taiwanese females with rheumatoid arthritis, primary Sjögren's syndrome and secondary Sjögren's syndrome in rheumatoid arthritis. Journal of Proteomics. 2016, 141:1-11.
Lin HI, Wu CC, Yang CH, Chang KW, Lee GB, Shiesh SC. Selection of aptamers specific for glycated hemoglobin and total hemoglobin using on-chip SELEX. Lab on a Chip. 2015, 15:486-494.
Marchetti P. Advanced glycation end products (AGEs) and their receptors (RAGEs) in diabetic vascular disease. Medicographia. 2009, 257-265.
Moon JM, Kim DM, Kim MH, Han JY, Jung DK, Shim YB. A disposable amperometric dual-sensor for the detection of hemoglobin and glycated hemoglobin in a finger prick blood sample. Biosensors and Bioelectronics. 2017, 91: 128-135.
Mozzarelli A, Bruno S, Ronda L. Hemoglobin-based oxygen carriers as red cell substitutes and oxygen therapeutics. Springer, New York. 2013, 55-73.
Nathan DM, Kuenen J, Borg R, Zheng H, Schoenfeld D, Heine RJ. Translating the A1C assay into estimated average glucose values. Diabetes Care. 2008, 31:1473-1478.
Sanavio B, Krol S. On the slow diffusion of point-of-care systems in therapeutic drug monitoring. Frontiers in Bioengineering and Biotechnology. 2015, 3:1-15.
Sebastian C, William H. Immunochromatography: formats and application. Indo American Journal of Pharmaceutical Research. 2016, 6400-6417.
Stark GR, Alwine JC, Kemp DJ. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proceedings of the National Academy of Sciences of the United States of America. 1977, 5350-5354.
Trier N, Houen. Epitope specificity of anti-citrullinated protein antibodies. Antibodies. 2017, 6.
Wang JP, Yoo SK, Song L, Mostafa A. El-Sayed. Molecular Mechanism of the Differential Photoelectric Response of Bacteriorhodopsin. Journal of Physical Chemistry B. 1997, 101:3420-3423.
Wang J. Vectorially oriented purple membrane: characterization by photocurrent measurement and polarized-Fourier transfor infrared spectroscopy. Thin Solid Films 2000, 379:224-229.
周文娟, 吳云翔, 易軍. 糖化血红蛋白標準化及其檢測技術的發展. 生物化學與生物物理進展.2015, 42: 443-456.
World Health Organization, Diabetes, 2013
https://web.archive.org/web/20130826174444/http://www.who.int/mediacentre/factsheets/fs312/en/
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) , Gestational Diabetes, 2014
https://www.niddk.nih.gov/health-information/diabetes/overview/what-is-diabetes/gestational?dkrd=hiscr0003
廖信銓. 微流體於細菌視紫質光電晶片製備之應用. 國立台灣科技大學化學工程研究所碩士論文. 2015.
鄭凱如. 細菌視紫質泛用型免疫光電晶片製備與原子力顯微鏡分析. 國立台灣科技大學化學工程研究所碩士論文. 2012.
郭力彬. 紫膜生物光電晶片應用於糖化血紅素(HbA1c)檢測之探討. 國立台灣科技大學化學工程研究所碩士論文. 2017.
鄭智文. 以紫膜生物光電晶片檢測鉛離子及糖化血紅素. 國立台灣科技大學化學工程研究所碩士論文. 2018.
王詠毅. 紫膜生物光電晶片之類風濕性關節炎檢測應用與園子利顯微鏡分析. 國立台灣科技大學化學工程研究所碩士論文. 2018.
龔庭萱. 新穎性血清蛋白質之丙二醛修飾在類風濕關節炎致病角色之探討. 台北醫學大學醫學檢驗暨生物技術研究所碩士論文. 2016.