中文摘要
退伍軍人菌及肺炎披衣菌是常見的醫院內及社區性肺炎的重要致病菌。在年長者或是加護病房病患感染則容易發生較高之致死率。目前使用於檢驗退伍軍人桿菌感染的方法缺乏足夠的敏感度,有時特異性亦有不足，在肺炎披衣菌方面亦是如此。因此本實驗建立一套實用的分子診斷方法來診斷退伍軍人菌及肺炎披衣菌。在退伍軍人菌方面，我們以16S ribosomal subunits及MIP gene當作目標基因，16S rRNA上面的特殊序列可以幫助我們辨識所有的退伍軍人菌屬，而MIP gene則對嗜肺性退伍軍人菌有高度專一性。在肺炎披衣菌方面我們以pst-1 fragment當作我們的目標序列，因該段序列對肺炎披衣菌具有高度專一性，在這裡我們以Real-time PCR作為我們診斷的工具，並以SYBR Green 1為試劑，配合melting curve的分析作為判斷之依據。整個2004年共有165位社區型肺炎診斷出來，其中134位有痰液的分泌。在退伍軍人菌方面我們診斷出5位Real-time PCR陽性但一般檢驗法為陰性的病患。在肺炎披衣菌方面也相對檢驗出6位血清抗體陰性但Real-time PCR陽性的病患。因此，我們認為Real-time PCR的確為臨床診斷相當有效率的工具，對於社區型肺炎的調查及控制都有助益。

Abstract
Legionella species and Chlamydia pneumoniae are a common cause of community-acquired pneumonia and occasional cause of hospital-acquired pneumonia. Significant mortality rates among the elderly and patients with severe underlying disease may occur as a result of infection with those pathogen. Diagnostic delay may also result in increased mortality. Therefore, nucleic acid amplification assays have been shown to be useful for the detection of Legionella.spp and Chlamydia pneumoniae. The genes that encode 16S ribosomal subunits and the macrophage infectivity potentiator (MIP) gene have been shown to contain signature sequences that are useful for the identification of L. pneumophila and a variety of other Legionella species. The pst-1 fragment is useful for identification of Chlamydia pneumoniae. Here we try to test clinical specimens by Real-time PCR assays to detect L.pneumophila and other Legionella species in the same tube, and detect Chlamydia pneumoniae by SYBR Green 1 reagent. By this method, these amplicons of 16S ribosomal subunits gene and MIP gene can be discriminated by different melting curve dependent on different Tm values. In this study, we detected more 5 and 6 patients in Legionella species and Chlamydia pneumoniae than conventional diagnostic tools. Hence, the Real-time PCR also demonstrated that it’s a rapid and high sensitivity method in diagnosis of legionnaires’ disease. In this study, it also demonstrated that Real-time PCR is effective in prediction of atypical pathogen infection.

目錄
中文摘要--------------------------------------------------Ⅳ
英文摘要--------------------------------------------------Ⅴ
導論-------------------------------------------------------1
臨床肺炎之簡介---------------------------------------------1
退伍軍人桿菌簡介-------------------------------------------2
退伍軍人桿菌之歷史介紹-------------------------------------3
退伍軍人症之微生物特性介紹---------------------------------4
嗜肺性退伍軍人桿菌治病機制---------------------------------5
退伍軍人症的傳染途徑---------------------------------------6
退伍軍人症之診斷方法---------------------------------------7
披衣菌簡介------------------------------------------------11
披衣菌的感染----------------------------------------------12
披衣菌之微生物特性及感染機制------------------------------13
臨床披衣菌之診斷方法--------------------------------------14
退伍軍人菌及披衣菌之分子偵測------------------------------16
即時聚合

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