光滑假丝酵母菌对氟康唑耐药的相关研究

发布时间：2018-05-19 21:42

本文选题：光滑假丝酵母菌 + 耐药　；参考：《安徽医科大学》2017年硕士论文

【摘要】：第一部分光滑假丝酵母菌的耐药分析与快速鉴定目的了解光滑假丝酵母菌流行病学情况,以及2014年-2016年三年间临床常用抗真菌药物的耐药性和敏感性,总结其在标本来源、临床科室及不同年龄的分布特点,归纳近年来其耐药率的变化,同时评价MALDI-TOF在快速鉴定光滑假丝酵母菌中的应用,为疾病监测和治疗提供参考。方法收集安徽医科大学第一附属医院2014至2016三年间所有分离出的假丝酵母菌的数据,使用Whonet5.6软件分析光滑假丝酵母菌临床和药敏资料;并用MALDI-TOF系统和ITS测序法分别对其中的光滑假丝酵母菌进行菌种验证,以ITS测序法作为金标准,比较两种方法结果准确性。结果光滑假丝酵母菌在所有假丝酵母菌中所占比例为26.04%,排名第二。经过MALDI-TOF与ITS测序验证,所测菌株均为光滑假丝酵母菌。经过两种方法鉴定,MALDI-TOF与ITS测序在鉴定光滑假丝酵母菌时结果准确率没有差异。感染光滑假丝酵母菌的患者中,60岁以上老人最多,占66.73%;重症医学科和呼吸内科菌株所占比例高,分别为28.32%和9.7%。2014-2016年间光滑假丝酵母菌对临床常用抗真菌药物耐药率有所上升,其中对氟康唑耐药率分别是18.4%、21.9%、22.3%。结论MALDI-TOF在鉴定光滑假丝酵母菌的准确率高,适合在临床实验室推广。光滑假丝酵母菌具有明显的临床分布特征,近三年对临床常用抗真菌药物耐药率有所上升。第二部分光滑假丝酵母菌对氟康唑耐药的研究目的探讨光滑假丝酵母菌PDR1基因突变与氟康唑耐药的关系。方法从第一部分鉴定出的光滑假丝酵母菌中随机选取13株,用微量肉汤稀释法验证各菌株的药敏结果。PCR反应扩增耐药菌株PDR1基因,产物经琼脂糖凝胶电泳后测序,通过BLAST网站比对序列,分析有无突变。结果经药敏验证氟康唑耐药和敏感株均存在,13株中有9株耐药,4株不耐药。4株非耐药株中没有发现错义突变株,氟康唑耐药的9株光滑假丝酵母菌中,有5株存在氨基酸突变。突变位点有4种,分别是L280F,T588A,Y584C和P822L。结论发现氟康唑耐药的光滑假丝酵母菌PDR1基因存在氨基酸突变,其突变位点有4种(L280F,T588A,Y584C和P822L),它们可能参与光滑假丝酵母菌对氟康唑的耐药。
[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.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R446.5

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