臺灣博碩士論文加值系統

中文摘要
Bacteroides fragilis group乃專一性厭氧菌中最常於臨床檢體分離出之一群革蘭氏陰性厭氧桿菌。至目前為止至少包含有九種species。
其為腸道正常菌叢但會伺機性感染人體造成陰道炎、腹膜炎、闌尾炎、膽囊炎等; 嚴重時會引發下腹部之菌血症或敗血症。其中以B. fragilis菌株臨床分離率最高，B. thetaiotaomicron次之，二者造成之感染亦最常見。此菌屬對藥物之抗性有逐漸增加之趨勢，且不同species間也有一些差異，而主要之傳統鑑定方法及藥物感受性試驗則相當耗時費力，故學者們多致力於快速鑑定之研究。
近年來利用分子生物學技術，一些廣泛存在於細菌體之基因已被應用於菌種鑑定。本實驗嘗試利用16S rRNA gene及heat shock protein
gene來鑑定常見之B. fragilis group : B. fragilis、B. distasonis、B. caccae B. thetaiotaomicron、B. ovatus、B. uniformis、B. vulgatus。
實驗首先以引子8FPL、1492放大菌種16S rRNA gene近全長之片段，再利用RsaΙ、HinfΙ、HaeIII三種限制酵素切割後跑電泳用以分型。其中主要之B. fragilis以RsaΙ、HaeIII; B. thetaiotaomicron以HaeIII切割分別可得到獨特之pattern。接著利用heat shock protein 60其中一段於多數菌體間相當普遍存在之相似序列( groEL conserved sequences )設計之degenerate primers來放大B. fragilis group七種ATCC標準菌株以進行核酸序列及菌種演化關係之分析，並分析臨床菌株之序列得著與標準菌株具98.7∼100﹪的相似性應可應用於鑑定之結果。並依此序列比對，找出其中差異性較大之處設計出B. fragilis菌株鑑定用之菌種特異性引子( species-specific primers )，次以55株B. fragilis 臨床菌株及其他30株厭氧、需氧菌測試，得到100 ﹪之敏感度及特異性。此外亦利用PCR方式偵測B. fragilis臨床菌株對數種常用藥物之抗藥基因，並結合B. fragilis菌種特異性引子及ermF( clindamycin抗藥基因)引子設計成複合式PCR 。 於24株clindamycin抗藥菌株中，有12株( 50﹪)可測得ermF基因;其中11株可同時測得兩種標的之特異性片段。此結果顯示B. fragilis對clindamycin之抗藥性除ermF外，應有其他機制參與其中。

目錄
英文摘要…………………………………………………………………..1
中文摘要…………………………………………………………………..3
緒論………………………………………………………………………..5
實驗設計架構……………………………………………………………..9
材料與方法………………………………………………………………10
結果………………………………………………………………………28
討論………………………………………………………………………33
實驗結果之附圖表………………………………………………………38
參考文獻…………………………………………………………………62

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