臺灣博碩士論文加值系統

多數的抗甲氧苯青黴素金黃色葡萄球菌(Methicillin-resistant Staphylococcus
aureus, MRSA)被歸屬於醫療照護相關之MRSA (Healthcare-associated MRSA,
HA-MRSA)的範疇，且被廣泛認為是院內感染(Nosocominal infection)主要致病原，
危及患者生命安全，而在醫院的環境中也常從皮膚或軟組織分離出社區型相關
MRSA (Community -associated MRSA, CA-MRSA)，相對於HA-MRSA，這類的金
黃色葡萄球菌通常會表現白血球毒素基因(Panton-valentine leukocidin, PVL)。
本研究中，我們在2007-2010 年期間，從來國軍高雄總醫院看診或住院病人的各
類檢體中分離並收集了320 株MRSA 菌株，除了透過一般例行性培養法
(Culture-based method)加以鑑定外，抗生素感受試驗(Antibiotics susceptibility test)
也一併篩檢。另一方面藉由多重分子聚合脢連鎖反應 (Multiplex polymerase chain
reaction, mt-PCR)的方法，也對菌株的抗藥基因分型 (Genotype)加以鑑定，而MRSA
的spa 分型結果也以DNA 直接定序(Direct sequencing)的方法作確認。
在本論文之研究所探討MRSA 菌株抗藥性，主要包括菌株的SCCmec 分型、有
無表現mecA 以及PVL 基因等。另外也評估上述的分子形質配合細菌流行病學資
料有否區分醫院型與社區型MRSA 菌群之能力以及這些資料可否預測本院之院內
感染。從SCCmec 分型的結果顯示，本院所分離的MRSA 大多帶有SCCmec type IV
及V 分型(66.6%)，且大部分具有多重抗藥性(Multiple-drug resistant, MDR)。本研
究利用多重分子聚合酶連鎖反應進行MRSA 抗藥基因多型性檢測，並建置基礎細
菌分子資料庫，期望未來做為臨床抗生素用藥選擇時的參考。

A majority of the methicillin-resistant Staphylococcus aureus (MRSA) was thought as
Healthcare-associated MRSA (HA-MRSA), which has been recognized as main
causative agent of nosocomial infection. Contrarily, the community-associated MRSA
(CA-MRSA) was reported that frequently isolated from skin and soft tissues in a
hospital environment. In addition, these MRSA isolates usually expressed PVL gene
(Panton-valentine leukocidin, PVL). In this study, we have collected 320 bacterial
isolates comprised of pre-identified MRSA isolates from the specimens of patients in
Kaohsiung Armed Forces General Hospital during 2007 to 2010. For bacterial
identification, in addition to routinely culture-based method, the antibiotics
susceptibility test was performed, and multiplex polymerase chain reaction (mt-PCR)
was implemented for SCCmec typing and each observed spa type was confirmed by
DNA direct sequencing. In this study, we investigated the drug resistance of those
collected MRSA isolates by which genotyping of SCCmec as well as detecting on
whether presecnce of mecA and PVL. Moreover, the clinical impact of whether
intergretiing molecular data and bacterial epidemiological information can successfully
distinguishs the CAMRSA from HA-MRSA and predicts nosocomial infection in our
hospital was evaluated. As a result, we found a majority of the MRSA isolates were
SCCmec IV or V (66.6%) with multiple-drug resistance (MDR). In summry, we utilize
mt-PCR technique for SCCmec genotyping and to establish the molecular database of
MRSA strains in this study. Our results might provide a quick reference for decision
making of antimicrobial therapy.

論文審定書…………………………………………………………………………… i
致謝…………………………………………………………………………………… ii
中文摘要………………………………………………………………………………iii
英文摘要………………………………………………………………………………iv
研究目的………………………………………………………………………………vi
前言： .............................................................................................................................. 1
第一章、抗藥性機制: ..................................................................................................... 1
1.1 青黴素結合蛋白(Penicillin-binding proteins2a；PBP2a): .............................. 1
1.2 葡萄球菌卡匣染色體(Staphylococcal chromosome cassette mec；SCCmec):
.................................................................................................................................. 2
1.3 白血球毒素基因(Panton-valentine leukocidin；PVL)： ................................ 3
1.4 葡萄球菌蛋白A 基因（Staphylococal protein A-encoding gene；Spa）： . 4
第二章、HA-MRSA 和CA-MRSA 的介紹: ................................................................. 4
2.1 醫療照護相關抗甲氧苯青黴素金黃色葡萄球菌(Healthcare-associated；
HA-MRSA)： .......................................................................................................... 4
2.2 社區相關抗甲氧苯青黴素金黃色葡萄球菌(Community-associated MRSA；
CA-MRSA)： ........................................................................................................... 5
第三章、材料與方法 ...................................................................................................... 6
3.1 臨床菌株收集與來源: ....................................................................................... 6
3.2 臨床菌株確認培養: ........................................................................................... 6
3.3 抗生素敏感性試驗(Antimicrobial susceptibility test): ..................................... 6
3.4 菌株的純化: ....................................................................................................... 7
3.5 PVL 基因檢測: ................................................................................................... 7
3.6 SCCmec 基因檢測: ............................................................................................ 8
3.7 Spa 基因檢測及分型: ........................................................................................ 8
3.8 DNA 直接定序(Direct sequencing): .................................................................. 9
3.9 統計分析: ........................................................................................................... 9
第四章、結果 ................................................................................................................. 11
4.1 收集菌株統計: .................................................................................................. 11
4.2 抗生素感受試驗(Antimicrobial susceptibility test): ........................................ 11
4.3 SCCmec 基因檢測: .......................................................................................... 12
4.4 PVL 基因檢測: ................................................................................................. 12
4.5 Spa 基因檢測及分型: ...................................................................................... 12
第五章、討論： ............................................................................................................ 13
參考文獻： .................................................................................................................... 17
圖表目錄： .................................................................................................................... 21

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