"表面增強拉曼光譜細菌藥敏測試法” 所利用的生物標記之分子來源探討

Molecular Origin of the Surface-Enhanced-Raman-Scattering Biomarkers Exploited for Bacterial Antibiotic Susceptibility Test

邱文瑜(Shirley Wen-Yu Chiu)

指導教授 ： 王玉麟 ； 楊家銘

國立清華大學/理學院/化學系所/博士(2020年)

摘要

表面增強式拉曼光譜由於其快速偵測及非侵入性的特性，已經被廣泛地應用在微生物領域的研究上。近年來，細菌表面增強式拉曼光譜中的生物標記也被成功地運用在細菌的抗生素藥物敏感測試。本論文的目標為探討細菌在表面增強拉曼光譜結合藥物敏感測試中的生物標記之分子來源及其所代表的生理意義。我們使用了表面增強拉曼光譜及超高效液相層析串聯電噴灑式質譜儀，鑑定了格蘭氏陽性菌金黃色葡萄球菌與格蘭氏陰性菌大腸桿菌之生物標記的分子來源為來自細菌代謝釋放之數個嘌呤衍生物，更利用質譜將細菌在飢餓壓力之下釋放的分子隨時間作定量分析
。除此之外，為了瞭解細菌釋放此嘌呤衍生物之緣由，我們分析了數個核糖核酸降解基因缺失的大腸桿菌細菌株的拉曼光譜及質譜，證實細菌釋放之嘌呤衍生物與飢餓誘發之核醣體核糖核酸降解之關聯性。

關鍵字

表面增強式拉曼光譜 ； 抗生素藥物敏感測試 ； 超高效液相層析串聯電噴灑式質譜 ； 嘌呤 ； 核糖體核糖核酸降解

並列摘要

Surface-enhanced Raman scattering (SERS), due to its rapid and non-invasive characteristics, has be widely used in microbiological fields. Recently, the SERS biomarkers from bacteria were also applied to bacterial antibiotic susceptibility testing (AST) successfully. The aim of this work is to explore the origin of the biomolecules responsible for the bacterial biomarkers in the SERS-AST method and the underlying physiological implication. We used both SERS and UPLC/ESI-MS to identify the molecules released from Gram-positive bacteria Staphylococcus aureus and Gram-negative bacteria Escherichia coli as several purine derivatives, as well as to quantify the time-dependence concentration of the purine derivatives from bacteria in starvation stress. Furthermore, changes in both the SERS and MS spectral features in Escherichia coli mutants without specific RNA degrading enzymes were observed, verifying that the origin of the purine derivatives from bacteria is attributed to the ribosomal RNA degradation upon starvation.

並列關鍵字

Surface-enhanced Raman Scattering ； SERS ； Antibiotic susceptibility test (AST) ； UPLC/ESI-MS ； purine metabolites ； ribosomal RNA degradation

參考文獻

1. C. Y. Liu, Y. Y. Han, P. H. Shih, W. N. Lian, H. H. Wang, C. H. Lin, P. R. Hsueh, J. K. Wang and Y. L. Wang, Sci Rep, 2016, 6, 15.

2. Y.-Y. Han, Y.-C. Lin, W.-C. Cheng, Y.-T. Lin, L.-J. Teng, J.-K. Wang and Y.-L. Wang, Sci Rep, 2020, 10, 12538.

3. C. V. Raman and K. S. Krishnan, Nature, 1928, 121, 501-502.

4. G. Keresztury, in Handbook of Vibrational Spectroscopy2006.

5. P. R. G. Derek J. Gariner, 1989.