2018-08-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/686085摘要：肺囊蟲(Pneumocystis jirovecii)為台灣常見的黴菌，在愛滋病及免疫不全的病患中會造成嚴重的肺囊蟲肺炎(Pneumocystis pneumonia, PCP)，其預防及治療皆需使用磺胺劑(trimethoprim-sulfamethoxazole)。過去我們在台灣成功建立real-time PCR檢驗平台，並發現如同其他全球的報告，發生在非愛滋病患者的肺囊蟲肺炎正快速上升，其病程進展更迅速，死亡率也更高(35-60%)。對磺胺劑抗藥及特定基因型之超強菌株可能是造成高死亡率的原因之一，過去的小型研究亦發現，具磺胺劑抗藥相關基因變異的菌株會提高致死率，而在非洲愛滋病患者的研究發現，基因變異的發生高達50%以上。然而，因為肺囊蟲無法在體外培養進行藥物敏感性試驗，而基因變異檢測並非常規檢驗，並無法提供嚴重感染或治療反應不佳而需調整處方病患即時性參考，往往在治療不理想後才使用第二線藥物進行救援治療，延誤有效治療。目前雖然台灣非愛滋病患者的肺囊蟲肺炎發生率快速上升，但對磺胺劑抗藥相關基因變異的發生率、危險因子仍缺乏詳細研究，對於致病菌株間的分子相關性以及臨床表現亦無研究。本試驗希望於台大醫院收集二年共400個以上肺囊蟲檢測陽性檢體，進行磺胺劑抗藥相關基因變異檢測，以了解基因變異發生率及與病患治療結果相關性。第一年除收集200個以上檢體，針對抗藥基因變異序列加以分析比較外，第二年將運用第一年所收集之資料及過去建立自動化 real-time PCR 檢驗平台的經驗，建立新型自動化抗藥快速分子診斷平台，並再收集200個以上獨立檢體，加以驗證該平台之可靠性。我們亦將利用3種分子分型，分析各菌株的分子相關性，臨床表現，並建立相關性資料庫，分析了解菌株之群聚性，以利後續快速分型並提供傳染散佈途徑的分析參考。此研究的結果，將有助於幫助臨床醫師給予正確的治療，辨認可能發生治療失敗之高危險群，改善肺囊蟲肺炎的治療預後<br> Abstract: Pneumocystis jiroveci pneumonia (PCP) is a life-threatening fungal infection in immunocompromised patients, especially those with AIDS. The incidence of PCP in HIV-infected patients has already declined but increasing PCP cases has been noted in recent 10 years among non-HIV infected population such as malignancy, hematology disorder and autoimmune disease. Severe PCP in the disease in non-HIV infected patients tends to progress more rapidly and is associated with higher mortality rates (35-60%) than in patients with HIV. The most commonly used drug for the treatment of PCP is a trimethoprim-sulfamethoxazole (TMP–SMX) combination and presence of associated mutation are associated with treatment failure. However, the prevalence of TMP–SMX resistance associated genes mutations, risk factor, clinical impact and molecular epidemiology are still unknown in non-HIV infected patients. In current study, we will determinate TMP–SMX resistance associated genes mutations of 400 isolates within 2 years. In the first year, we will evaluate the prevalence and associated risk factors of determinate TMP–SMX resistance associated genes mutations in more than 200 RT-PCR+ specimens. In the secondary year, the mutation codons distribution will be used to develop an automatically molecular testing platform which will be validated with more than 200 specimens. Besides, Genetic relatedness will be performed by using 3 different genotyping method. The results of genes mutations, genotyping and clinical outcome would be clarified to seek potentially rapid diagnostic methods for identifying drug resistant or virulent strains from clinical isolates. The results will help to establish early and rapid diagnosis, as well as, appropriate treatment strategy to improve outcomes.肺囊蟲複方磺胺類抗生素抗藥Pneumocystis jiroveciitrimethoprim-sulfamethoxazoledrug resistance