- 學位論文
電漿應用於術後金黃色葡萄球菌之殺菌效果探討
The effect for inactivation of Staphylococcus aureus in postoperative by Plasma
摘要
本研究主軸為氦氣電漿對金黃色葡萄球菌之滅菌效率，相較傳統抗生素藥物療法之探討。研究大綱建構於三項主體，其一為自製之噴射式常壓氦氣電漿設備（Atmospheric-Pressure Glow Discharge Helium Plasma Jet Device），再者為醫學臨床上常引發傷口感染兩株病菌-金黃色葡萄球菌 ( Staphylococcus aureus, OSSA )、抗藥性金黃色葡萄球菌 ( Oxacillin-Resistant Staphylococcus aureus, ORSA )，以及選用苯唑西林 ( Oxacillin ) 抗生素藥品，藉由以上主體進行本研究規劃。 本研究首先欲驗證OSSA、ORSA之藥物敏感性，備製苯唑西林 2、4 μg/ml兩種濃度，各別與兩菌株共培養0、4、8 hrs，檢測在相同條件下的存活率，結果分析顯示苯唑西林與OSSA共培養4、8 hrs，兩者皆達到99 %抑菌效果；ORSA呈現一概的抗藥結果。接續進行菌株施打電漿的驗證，展現電漿作用於OSSA、ORSA個別僅需20 mins即具備100 %滅菌的結果，顯示電漿殺菌效率相較抗生素更為顯著；研究第三階段進行電漿施打後再加入苯唑西林4 μg/ml共培養 10 mins，觀察其結果發現，電漿加抗生素實驗組，相較單獨施打電漿組，滅菌效率更提高，最後以電子顯微鏡觀察細菌表面，發現電漿處理後病菌形貌產生皺縮及變形，證明電漿的確在細菌表面產生蝕刻等作用，造成細菌失活。 本研究之價值在於證明電漿於滅菌治療上，已具備針對抗藥性之微生物，提供一種有效成功的滅菌方式，可作為替代傳統抗生素傷口抗菌的另一選擇，其低溫、快速有效滅菌之優點，有別於傳統藥物治療的種種副作用與限制，值得爾後深入探討，擴展其對於多重性抗藥病菌之應用性。 關鍵詞：噴射式氦氣低溫電漿、抗生素、金黃色葡萄球菌、抗藥性
並列摘要
This study was conducted to compare the in vitro effectiveness of in-house developed Plasma, that is produced by the Atmospheric-Pressure Glow Discharge Helium Jet Plasma Device on bacterial sterilization, versus antibiotic therapy. Staphylococcus aureus is a common bacterial pathogen found in wound infection and can be treated with the antibiotic Oxacillin with various degrees of success. The effectiveness of Oxacillin is diminished in Oxacillin-resistant S. aureus (ORSA) versus Oxacillin-sensitive S. aureus (OSSA), limiting the value of antibiotic therapy in cases of antibiotic-resistant strains. In the first stage of the study, ORSA and OSSA were grown out in medium and exposed to two concentrations of Oxacillin (2 μg/ml and 4 μg/ml). Survival was assessed at 0, 4 and 8 hours. Results demonstrated sensitivity and resistance to Oxacillin by OSSA and ORSA, respectively, as expected. In the second stage of the study, the two bacterial strains were exposed to Plasma for 5, 7, 10, 15, and 20 minute time intervals. A separate group was exposed to Plasma for 5, 7 and 10 minute time intervals and then co-cultured with Oxacillin at 4 μg/ml concentration. Results showed a greater sterilization efficacy of Plasma versus Oxacillin. Specifically, exposure of both OSSA and ORSA to Plasma for 20 minutes resulted in 100 % sterilization. Results also showed greater effectiveness when Plasma was combined with Oxacillin versus single modality treatment with Oxacillin 4 μg/ml alone or Plasma 10 minutes alone. In the third stage of the study, sterilization by Plasma was confirmed by FE-SEM (field emission scanning electron microscopy), which demonstrated shrinkage and deformation of bacteria following Plasma treatment. This study demonstrates the in vitro effectiveness of Plasma treatment as a viable option for sterilization of Oxacillin-sensitive and Oxacillin-resistant strains of S.aureus. Plasma may be a promising alternative to antibiotic treatment of infected wounds in the clinical setting. It provides low temperature, rapid-effectiveness sterilization and as such, it may overcome or complement traditional antibiotic therapy which has its own shortcomings due to prevalence of antibiotic resistance. Consequently, Plasma deserves further research in order to expand its applications in combating multidrug-resistant pathogenic bacteria. Key words: Oxacillin, OSSA, ORSA, MRSA, Plasma