本文選題：光滑假絲酵母菌 + 耐藥��； 參考：《安徽醫(yī)科大學(xué)》2017年碩士論文

【摘要】：第一部分光滑假絲酵母菌的耐藥分析與快速鑒定目的了解光滑假絲酵母菌流行病學(xué)情況,以及2014年-2016年三年間臨床常用抗真菌藥物的耐藥性和敏感性,總結(jié)其在標(biāo)本來(lái)源、臨床科室及不同年齡的分布特點(diǎn),歸納近年來(lái)其耐藥率的變化,同時(shí)評(píng)價(jià)MALDI-TOF在快速鑒定光滑假絲酵母菌中的應(yīng)用,為疾病監(jiān)測(cè)和治療提供參考。方法收集安徽醫(yī)科大學(xué)第一附屬醫(yī)院2014至2016三年間所有分離出的假絲酵母菌的數(shù)據(jù),使用Whonet5.6軟件分析光滑假絲酵母菌臨床和藥敏資料;并用MALDI-TOF系統(tǒng)和ITS測(cè)序法分別對(duì)其中的光滑假絲酵母菌進(jìn)行菌種驗(yàn)證,以ITS測(cè)序法作為金標(biāo)準(zhǔn),比較兩種方法結(jié)果準(zhǔn)確性。結(jié)果光滑假絲酵母菌在所有假絲酵母菌中所占比例為26.04%,排名第二。經(jīng)過(guò)MALDI-TOF與ITS測(cè)序驗(yàn)證,所測(cè)菌株均為光滑假絲酵母菌。經(jīng)過(guò)兩種方法鑒定,MALDI-TOF與ITS測(cè)序在鑒定光滑假絲酵母菌時(shí)結(jié)果準(zhǔn)確率沒(méi)有差異。感染光滑假絲酵母菌的患者中,60歲以上老人最多,占66.73%;重癥醫(yī)學(xué)科和呼吸內(nèi)科菌株所占比例高,分別為28.32%和9.7%。2014-2016年間光滑假絲酵母菌對(duì)臨床常用抗真菌藥物耐藥率有所上升,其中對(duì)氟康唑耐藥率分別是18.4%、21.9%、22.3%。結(jié)論MALDI-TOF在鑒定光滑假絲酵母菌的準(zhǔn)確率高,適合在臨床實(shí)驗(yàn)室推廣。光滑假絲酵母菌具有明顯的臨床分布特征,近三年對(duì)臨床常用抗真菌藥物耐藥率有所上升。第二部分光滑假絲酵母菌對(duì)氟康唑耐藥的研究目的探討光滑假絲酵母菌PDR1基因突變與氟康唑耐藥的關(guān)系。方法從第一部分鑒定出的光滑假絲酵母菌中隨機(jī)選取13株,用微量肉湯稀釋法驗(yàn)證各菌株的藥敏結(jié)果。PCR反應(yīng)擴(kuò)增耐藥菌株P(guān)DR1基因,產(chǎn)物經(jīng)瓊脂糖凝膠電泳后測(cè)序,通過(guò)BLAST網(wǎng)站比對(duì)序列,分析有無(wú)突變。結(jié)果經(jīng)藥敏驗(yàn)證氟康唑耐藥和敏感株均存在,13株中有9株耐藥,4株不耐藥。4株非耐藥株中沒(méi)有發(fā)現(xiàn)錯(cuò)義突變株,氟康唑耐藥的9株光滑假絲酵母菌中,有5株存在氨基酸突變。突變位點(diǎn)有4種,分別是L280F,T588A,Y584C和P822L。結(jié)論發(fā)現(xiàn)氟康唑耐藥的光滑假絲酵母菌PDR1基因存在氨基酸突變,其突變位點(diǎn)有4種(L280F,T588A,Y584C和P822L),它們可能參與光滑假絲酵母菌對(duì)氟康唑的耐藥。
[Abstract]:The first part: drug resistance analysis and rapid identification of Candida smoothing objective to understand the epidemiological situation of Candida smooth candida and the drug resistance and sensitivity of common antifungal drugs in clinical use from 2014 to 2016, and to summarize its origin in specimens. The distribution characteristics of clinical departments and different ages were summarized, the changes of drug resistance rate in recent years were summarized, and the application of MALDI-TOF in rapid identification of Candida smoothing was evaluated to provide reference for disease surveillance and treatment. Methods the data of all Candida cerevisiae isolated from 2014 to 2016 in the first affiliated Hospital of Anhui Medical University were collected and the clinical and drug sensitive data of Candida smoothing were analyzed by Whonet5.6 software. MALDI-TOF system and ITS sequencing method were used to verify the species of Candida smoothing. ITS sequencing method was used as the gold standard to compare the accuracy of the two methods. Results the percentage of Candida smooth in all Candida cerevisiae was 26.04, ranking second. The results of MALDI-TOF and ITS sequencing showed that the strains were Candida smooth. There was no difference in accuracy between MALDI-TOF and ITS sequencing in the identification of Candida smooth. Most of the patients infected with Candida smooth were over 60 years old, accounting for 66.730.The proportion of strains in intensive medicine department and respiratory department were 28.32% and 9.73.The resistance rate of Candida smooth to common antifungal drugs increased between 2014 and 2016. The resistance rate to fluconazole was 18.4% and 21.9%, respectively. Conclusion MALDI-TOF has high accuracy in identification of Candida smoothing, and is suitable for clinical laboratory application. Candida cerevisiae has obvious clinical distribution, and the resistance rate to common antifungal drugs has increased in recent three years. The second part is the study of fluconazole resistance of Candida smoothing. Objective to investigate the relationship between fluconazole resistance and mutation of PDR1 gene in Candida smoothing. Methods Thirteen strains of Candida smooth were randomly selected from the first part of the study. The drug sensitivity of each strain was verified by broth dilution method. The PDR1 gene of resistant strain was amplified by PCR, and the product was sequenced by agarose gel electrophoresis. Through BLAST site alignment sequence, analysis of mutation. Results No missense mutant was found in 9 of 13 strains of fluconazole resistance and susceptible strains, and 5 of 9 strains of fluconazole-resistant Candida cerevisiae were found to have amino acid mutations. There are four mutation loci, L280FU T588AZY584C and P822L. Conclusion Amino acid mutations were found in the PDR1 gene of fluconazole-resistant Candida smoothing, and there were four kinds of L280FN T588AHY584C and P822L, which may be involved in the resistance of Candida smooth to fluconazole.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R446.5

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