| Summary: | 碩士 === 慈濟大學 === 醫學檢驗生物技術學系醫學生物技術碩士班 === 100 === PartΙ
The Multidrug-resistant strains of Acinetobacter baumannii (MDRAB) has increasingly been reported its serious problems worldwide beyond our expectations. Bacteriophage therapy is a potential alternative treatment for MDR bacterial infections. To study the multidrug resistance A. baumannii, we used mass spectrometry-based methods to analyze the protein profiles of A. bumannii bacteriophage. We collected ten force strains of A. baumannii phage from waste water of hospitals, following cultured and amplified using MDRAB. The phage proteins were extracted by water vortex and cleaned up by filtration and ultra-centrifuge, then proceeded by trypsin digested into peptides. A tip C18-column were used for the liquid chromatography to separate the peptide mixtures and detected by tandem MS under data-dependent mode. We Used BioworkTMsearch engine to search whole A. baumannii phage database to analyze the phage proteins, and predicted its function by bioinformatics tool. There are several proteins with similar predicted functions have been identified from different phage strains in this study. We have succeeded in clustering the A. bumannii phage with MS method. Additionally, one of them designed as ΦAB3, was further characterized. Electron microscopy showed that ΦAB3 was morphologically different in two types. Furthermore, based on the proposed proteomic approach, our study would provide bioinformation of bacteriophage a new insight into virus-host interaction and gene transgenosis in the future.
Part Π
Tuberculosis is one kind of high mortality rate human infectious disease caused by Mycobacterium tuberculosis, sometimes nontuberculous mycobacteria (NTM) confused the diagnosis of tuberculosis, and it’s difficult to make a difference in clinical manifestations and diagnostic evaluation. Therefore, the identification of Mycobacterium tuberculosis infection and nontuberculous mycobacteria is important. However, M. tuberculosis culture and identification need full 12 weeks which is incredible time-consuming procedure takes the laboratory cost. Besides, the percentage of resistant or multi-resistant of clinical strains have been increased in recent years. This phenomenon will threaten the ability of antibiotics to treat tuberculosis. To improve diagnostic methods and achieve fast and accurate results, rapid identification of Mycobacterium tuberculosis method is required. In this study, we use matrix-assisted laser desorption/ionization time of flight mass spectrometer (MALDI-TOF MS) for analysis. This method provides specific mass fingerprinting of Mycobacterium cell, and combine with the establishment of mass spectral database. It will be possible to rapidly identify clinical Mycobacterium species.
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