開發一套新型多型樣匹配方法結合基質輔助雷射脫附游離飛行時間質譜法用於提升細菌鑑定的可靠程度

Development of A New Multiple Pattern Matching Method for Robust Bacterial Identification by MALDI-TOF MS

林君叡(Chun-Jui Lin)

指導教授 ： 吳志哲

國立暨南國際大學/科技學院/應用化學學系/碩士(2021年)

https://doi.org/10.6837/ncnu202100225

摘要

近年來耐藥性致病細菌感染對全球公共衛生安全產生嚴重的威脅，因此臨床醫學上需要開發出一套能精準且快速的針對致病性細菌進行辨識的有效方法，才能對患者進行精準的治療。但傳統細菌鑑定的方式繁瑣且複雜，因此近年來大多使用細菌分子型(基因型鑑定及蛋白質鑑定)的鑑定方式，來達到快速且準確地鑑定。
本論文開發一套新型多型樣匹配(Multiple Pattern Matching)方法結合細菌質譜分析技術，能成功地提升菌種鑑定的可靠程度。首先，我們將細菌菌株加入醋酸鈉緩衝溶液(Sodium acetate buffer)進行裂解，利用不同有機溶劑對樣品中的脂質A和蛋白質進行萃取，再通過MALDI-TOF MS進行質譜分析來獲得細菌質譜圖。先將兩種不同的細菌質譜圖，利用細菌指紋質譜資料庫(Biotyper)軟體進行型樣匹配(Pattern Matching)演算法比對，可以分別獲得相對應的分數。再將獲得的分數進行平均，來獲得一個新的多重型樣匹配指標(MPM score)，這個新的指標可以幫我們提升細菌鑑定的可靠程度。這套新方法可以在1小時內完成細菌鑑定，與單一型樣匹配方法的相比，不僅可以直接從同一株細菌樣品中同時取得兩種不同的生物指紋資訊外，還可以應用再多種細菌混合物的鑑定上甚至尿液中的細菌混合物，不須透過細菌塗盤培養的複雜前處理步驟，且能更準確、高效的識別出複雜環境中的細菌菌種。在未來，我們會逐步擴大單株細菌的生物指紋資料庫，除了可以提升我們對複雜條件下細菌鑑定的準確度外，透過添加colistin-抗藥性的生物指紋特徵訊號，也能將這套方法成功地應用於識別colistin-抗藥性細菌菌種。

關鍵字

基質輔助雷射脫附游離飛行時間質譜 ； 多型樣匹配方法 ； 細菌鑑定

並列摘要

In recent years, drug-resistant pathogenic bacterial infections have posed a serious threat to global public health security. Therefore, clinical medicine needs to develop an effective method that can accurately and quickly identify pathogenic bacteria in order to treat patients accurately. However, the traditional identification method for bacterial characterization is time-consuming and laborious. Hence, most of them use bacterial molecular type (genotype identification and protein identification) identification methods to achieve rapid and accurate identification.
In this thesis, a new multiple pattern matching method combined with bacterial mass spectrometry analysis technology is developed, which can successfully improve the reliability of bacterial species identification. This new method can successfully identify bacteria within 1 hour. Compared with the single pattern matching method, it not only can directly obtain two different biological information from the same sample, but also in multiple mixtures or urine.In the future, we will gradually expand the biofingerprint database of individual bacteria. In addition to improving the accuracy of bacterial identification under complex conditions and colistin-resistant bacterial strains.

並列關鍵字

MALDI-TOF MS ； multiple pattern matching method ； bacterial identification

參考文獻

參考文獻

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