白念珠菌唑类耐药和生物膜相关基因的表达  

The expression of genes related to azole resistance and biofilm in Candida albicans

作　　者：胡绿荫[1] 宰淑蓓[1] 金鑫[1] 蔡金凤[1] 曹宇硕 胡香南[1] 杜昕[2] 李天铭[2] 李敏[3] 

机构地区：[1]上海市(复旦大学附属)公共卫生临床中心医学检验科,上海201508 [2]复旦大学附属华山医院检验科,上海200040 [3]上海交通大学医学院附属仁济医院检验科,上海200127

出　　处：《检验医学》2014年第6期597-602,共6页Laboratory Medicine

基　　金：国家自然科学基金资助项目(81171623;81261120387;81322025;81371875);上海优秀青年人才培养计划项目(XYQ2011039);上海曙光人才计划(12SG03)

摘　　要：目的研究白念珠菌临床分离株唑类耐药和生物膜形成相关基因的表达。方法收集上海市公共卫生临床中心2006至2011年92例感染性疾病患者[病毒性肝炎、结核、获得性免疫缺陷综合征(AIDS)]血液、无菌部位和黏膜分离鉴定的白念珠菌104株,选用ATB FUNGUS3检测抗菌药物(氟康唑、伊曲康唑和伏立康唑)的最低抑菌浓度(MIC),采用半定量逆转录聚合酶链反应(PCR)分析唑类敏感、浓度依赖性敏感(S-DD)和耐药菌株之间药物外排泵基因CDR1、CDR2、MDR1和药物靶酶基因ERG11表达的差异,同时通过甲基四氮盐(XTT)代谢试验筛选强生物膜形成能力菌株(HBFs),研究其与弱生物膜形成能力菌株(LBFs)相关基因ALS3和HWP1表达的差异。结果 104株白念珠菌中共检测出16株至少对1种唑类药物耐药,6株至少对1种唑类药物S-DD。药物靶酶基因ERG11在耐药株、S-DD株和敏感菌之间比较,差异有统计学意义(P=0.007 8),且耐药株、S-DD株分别与敏感菌比较,差异有统计学意义(P<0.05),然而CDR1、CDR2和MDR1表达量在3种菌株间无明显差异。生物膜形成能力试验显示16株菌生物膜形成能力增强,其HWP1表达量与LBFs比较差异有统计学意义(P=0.007 9),ALS3表达量差异无统计学意义。结论 ERG11基因的过度表达为白念珠菌唑类耐药的重要机制。菌丝细胞壁蛋白HWP1基因的高表达与白念珠菌生物膜形成增强相关。Objective To investigate the expression of genes related to azole resistance and biofilm formation capability in clinical isolates of Candida albicans.Methods In this study,1 04 Candida albicans were isolated from blood,sterile sites and mucosal lesions of 92 infectious disease patients [viral hepatitis,tuberculosis and acquired immune deficiency syndrome （AIDS）in Shanghai Public Health Clinical Center from 2006 to 201 1 .The minimum inhibitory concentrations （MICs）of fluconazole,itraconazole and voriconazole were determined by ATB FUNGUS3.The gene expression of efflux pump genes （CDR1 ,CDR2 and MDR1 ） and azole antifungal target gene ERG1 1 were determined by semi-quantitative reverse transcription polymerase chain reaction （PCR）,and the azole susceptibe and susceptible dose dependent （S-DD）states were analyzed.2,3-bis-（2-methoxy-4-nitro-5-sulphenyl）-2H-tetrazolium-5-carboxanilide （XTT）test was used to screen the high biofilm formation capability isolates （HBPs）.The expressions of ALS3 and HWP1 in low biofilm formation capability isolates （LBPs）were analyzed.Results Among 1 04 isolates of Candida albicans,1 6 isolates were tested resistant to at least one azole,6 isolates were S-DD to at least one azole.There were statistical significance among resistant,S-DD and susceptible isolates in azole antifungal target gene ERG1 1 （P=0.007 8）,and significant difference was found when comparing susceptible isolates with resistant and S-DD ones （P＆lt;0.05）.No significant difference was found in the expression levels of efflux pump genes,CDR1 ,CDR2 and MDR1 . The expression levels of HWP1 in HBFs were higher than those in LBFs with statistical significance （P=0.007 9 ）, whereas ALS3 showed no statistical significance.Conclusions The overexpression levels of ERG1 1 gene may primarily account for resistance to azole in Candida albicans.High expression of hyphal wall protein HWP1 gene is associated to＆amp;nbsp;biofilm formation capability in Candida albicans.

关 键 词：外排泵 ERG11 生物膜 白念珠菌 

分 类 号：R446.5[医药卫生—诊断学]