本文關(guān)鍵詞：呼吸道感染中白假絲酵母菌細(xì)胞外水解酶和耐藥性對菌株毒力的影響　出處：《福建醫(yī)科大學(xué)》2007年碩士論文　論文類型：學(xué)位論文

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【摘要】： 目的分析呼吸道白假絲酵母菌細(xì)胞外磷脂酶B和分泌型天冬氨酸蛋白酶活力及其基因表達(dá)對菌株毒力的影響;分析呼吸道白假絲酵母菌對唑類抗真菌藥物的耐藥情況與耐藥基因的表達(dá)關(guān)系;分析白假絲酵母菌毒力與耐藥性的關(guān)系。 方法①收集臨床呼吸道白假絲酵母菌菌株,通過培養(yǎng)鑒定將菌株分為致病組和非致病組;②采用卵黃培養(yǎng)基法檢測菌株細(xì)胞外磷脂酶B的活力,用牛血清白蛋白瓊脂培養(yǎng)基法檢測分泌型天冬氨酸蛋白酶的活力,分析組間差別;③用NCCLS M27-A方案中的微量液基稀釋法檢測菌株對氟康唑和伊曲康唑的最小抑菌濃度(MIC),比較菌株組間的耐藥性差別;④用半定量RT-PCR的方法檢測細(xì)胞外磷脂酶B和分泌型天冬氨酸蛋白酶相關(guān)基因PLB1和SAP2,以及耐藥相關(guān)基因CDR1和MDR1的mRNA表達(dá),分析菌株的毒力和耐藥性與相關(guān)基因表達(dá)以及耐藥性與耐藥基因的關(guān)系;⑤對CDR1、MDR1、PLB1和SAP2的基因表達(dá)進(jìn)行相關(guān)分析。 結(jié)果共收集了84株呼吸道臨床菌株,非致病組有28株,致病組有56株;致病組分泌型天冬氨酸蛋白酶的活力要高于非致病組,細(xì)胞外磷脂酶活力未見差別;致病組菌株的PLB1和SAP2的mRNA的量均高于非致病組;二組菌株之間,氟康唑和伊曲康唑的耐藥性和耐藥相關(guān)基因的轉(zhuǎn)錄無差別;呼吸道白假絲酵母菌對伊曲康唑的耐藥率要高于氟康唑;MDR1與PLB1、MDR1與SAP2、PLB1和SAP2基因在mRNA的水平上有明顯的正相關(guān)。 結(jié)論分泌型天冬氨酸蛋白酶活力是呼吸道白假絲酵母菌重要的毒力因子,PLB1和SAP2的mRNA轉(zhuǎn)錄水平的上調(diào)可能影響菌株的毒力,呼吸道菌株對伊曲康唑的耐藥率要高于氟康唑,菌株的毒力與耐藥性和耐藥相關(guān)基因的轉(zhuǎn)錄無關(guān);MDR1、PLB1和SAP2基因調(diào)控在mRNA的水平可能有相關(guān)性。
[Abstract]:Objective to analyze the effects of extracellular phospholipase B and secretory aspartate protease activity and gene expression on virulence of Candida albicans. To analyze the relationship between the resistance of Candida albicans to antifungal agents and the expression of resistance genes. The relationship between virulence and drug resistance of Candida albicans was analyzed. Methods 1 strains of Candida albicans in clinical respiratory tract were collected and identified by culture. The strains were divided into pathogenic group and non-pathogenic group. 2the activity of extracellular phospholipase B was detected by yolk medium method, and the activity of secretory aspartic protease was detected by bovine serum albumin Agar medium. 3 the minimal inhibitory concentration of fluconazole and itraconazole was detected by microdilution method in NCCLS M27-A scheme, and the drug resistance of strains was compared. 4 the extracellular phospholipase B and secretory aspartic protease related gene PLB1 and SAP2 were detected by semi-quantitative RT-PCR. The relationship between virulence and drug resistance and the expression of related genes and the relationship between drug resistance and drug resistance genes were analyzed. 5 correlation analysis of the gene expression of CDR1, MDR1, PLB1 and SAP2. Results A total of 84 clinical strains of respiratory tract were collected. There were 28 strains in non-pathogenic group and 56 strains in pathogenicity group. The activity of secretory aspartate protease in pathogenic group was higher than that in non-pathogenic group, but there was no difference in extracellular phospholipase activity. The amount of mRNA of PLB1 and SAP2 in pathogenic group was higher than that in non-pathogenic group. There was no difference in drug resistance and transcription between fluconazole and itraconazole. The resistance rate of Candida albicans to itraconazole was higher than fluconazole. There was a significant positive correlation between MDR1 and PLB1MDR1 and SAP2PLB1 and SAP2 genes at the mRNA level. Conclusion the activity of secretory aspartate protease is an important virulence factor of Candida albicans. The up-regulation of mRNA transcription level of PLB1 and SAP2 may affect the virulence of the strain. The resistance rate of respiratory tract strains to itraconazole was higher than that of fluconazole, and the virulence of the strains was not related to drug resistance and transcription of drug-resistance related genes. The regulation of MDR1 PLB1 and SAP2 gene may be related to the level of mRNA.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R378

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本文編號(hào)：1391279