Study On The Fluconazole Resistance Of Candida Glabrata

PART Ⅰ Analysis of drugs resistance and rapid identification of Candida glabrataObjective This research was to understand the epidemiology of Candida glabrata and the resistance and sensitivity of antifungal drug used in clinic commonly from 2014 to 2016 yeares.This aim of study was not only to summarize the distribution characteristics of samples source,clinical departments and age,but also explore the change in drug resistance rate andevaluate the ability of MALDI-TOF in rapid identification in order to provide some information for the monitoring and treatment of disease.Methods All the data of isolated Candida was collected in the first affiliated hospital of Anhui medical university from 2014 to 2016.The strains of Candida glabrata were onfirmed by the MALDI-TOF system and ITS sequencing method,respectively.Whonet 5.6 software was used to analyze retrospective clinic datas and drug sensitive information.Results The proportion of Candida glabrata in all Candida was 26.04%,ranking in the second.Both MALDI-TOF and ITS sequencing verified that the tested strains were Candida glabrata.There was no difference between MALDI-TOF and sequencing results.Among the patients infected with Candida glabrata,60 years of age and older accounted for the most,accounting for 66.73%.The proportion of severe medicine and respiratory medicine accounted for a high proportion of 28.32% and 9.7%,respectively.During the period of 2014-2016,the drug resistance rate of Candida glabrata was increased to some extent,and the resistance rate to fluconazole was 18.4%,21.9% and 22.3%,respectively.Conclusion The accuracy of MALDI-TOF used in the identification of Candida glabrata was high.The Candida glabrata had obvious clinical characteristic of distribution.In the past three years,the rate of antifungal drug resistance was to rise in some extentPART Ⅱ Study on the resistance of Candida glabrata t Fluconazole oObjective This study was performed to investigate the relationship between mutations in PDR1 gene of Candida glabrata and fluconazole resistanceMethods Thirteen strains of Candida glabrata were randomly selected from the first part.ITS sequencing method was used to identify the Candida glabrata.The method of 96-well plate microdilution was used to verify resistance of Candida glabrata to fluconazole.PDR1 gene in Candida glabrata with fluconazole resistance were amplified by PCR and sequenced after agarose gel electrophoresis products.Website of BLAST was used to analyze the sequence and find out mutations of PDR1.Results Four strains of Candida glabrata were susceptible-does dependent(SDD)to fluconazole and 9 were resistant.No amino acid mutation was found in SDD strains.Four kinds of amino acid mutation sites were found in fluconazole-resistant strains,namely L280 F,T588A,Y584 C and P822 L.Conclusion Four kinds of mutations exist in PDR1 gene of Fluconazole-resistant Candida glabrata.They are L280 F,T588A,Y584 C and P822 L,and may involved in the mechanism resistance to fluconazole.