国际医药卫生导报
國際醫藥衛生導報
국제의약위생도보
INTERNATIONAL MEDICINE & HEALTH GUIDANCE NEWS
2014年
16期
2417-2421
,共5页
王一如%冯文莉%杨静%奚志琴%乔祖莎
王一如%馮文莉%楊靜%奚誌琴%喬祖莎
왕일여%풍문리%양정%해지금%교조사
白念珠菌%ERG11基因%耐药性%基因突变%高表达
白唸珠菌%ERG11基因%耐藥性%基因突變%高錶達
백념주균%ERG11기인%내약성%기인돌변%고표체
Candida albicans%ERG11 gene%Resistance%Gene mutation%Over-expression
目的 探讨白念珠菌ERG11基因突变、高表达及二者的相互作用在氟康唑耐药中的作用.方法 采用PCR法扩增实验菌株的ERG11基因,并进行测序及生物信息学分析,筛查突变位点;抽提实验菌株总RNA,并逆转录合成cDNA,采用实时荧光定量PCR(FQ-RT-PCR)法检测ERG11基因的表达水平.结果 24株临床菌株中,共检测出ERG 11基因存在24个同义突变位点和5个错义突变位点,其中,E174A、T123I、V234F为新发位点;白念珠菌ERG11基因在耐药组中的表达量高于敏感组,差异具有统计学意义未发生错义突变的白念珠菌耐药株ERG11基因的表达量高于发生错义突变的,差异具有统计学意义.结论 白念珠菌ERG 11基因的5个错义突变A114V、E174A、T123I、Y132H、V234F可能与氟康唑耐药形成有关;ERG11基因的高表达可能与氟康唑耐药有关;ERG11基因的错义突变可能对该基因的高表达存在抑制作用.
目的 探討白唸珠菌ERG11基因突變、高錶達及二者的相互作用在氟康唑耐藥中的作用.方法 採用PCR法擴增實驗菌株的ERG11基因,併進行測序及生物信息學分析,篩查突變位點;抽提實驗菌株總RNA,併逆轉錄閤成cDNA,採用實時熒光定量PCR(FQ-RT-PCR)法檢測ERG11基因的錶達水平.結果 24株臨床菌株中,共檢測齣ERG 11基因存在24箇同義突變位點和5箇錯義突變位點,其中,E174A、T123I、V234F為新髮位點;白唸珠菌ERG11基因在耐藥組中的錶達量高于敏感組,差異具有統計學意義未髮生錯義突變的白唸珠菌耐藥株ERG11基因的錶達量高于髮生錯義突變的,差異具有統計學意義.結論 白唸珠菌ERG 11基因的5箇錯義突變A114V、E174A、T123I、Y132H、V234F可能與氟康唑耐藥形成有關;ERG11基因的高錶達可能與氟康唑耐藥有關;ERG11基因的錯義突變可能對該基因的高錶達存在抑製作用.
목적 탐토백념주균ERG11기인돌변、고표체급이자적상호작용재불강서내약중적작용.방법 채용PCR법확증실험균주적ERG11기인,병진행측서급생물신식학분석,사사돌변위점;추제실험균주총RNA,병역전록합성cDNA,채용실시형광정량PCR(FQ-RT-PCR)법검측ERG11기인적표체수평.결과 24주림상균주중,공검측출ERG 11기인존재24개동의돌변위점화5개착의돌변위점,기중,E174A、T123I、V234F위신발위점;백념주균ERG11기인재내약조중적표체량고우민감조,차이구유통계학의의미발생착의돌변적백념주균내약주ERG11기인적표체량고우발생착의돌변적,차이구유통계학의의.결론 백념주균ERG 11기인적5개착의돌변A114V、E174A、T123I、Y132H、V234F가능여불강서내약형성유관;ERG11기인적고표체가능여불강서내약유관;ERG11기인적착의돌변가능대해기인적고표체존재억제작용.
Objective To explore ERG11 gene mutations,over-expression and the relation between each other of Candida albicans in the role of fluconazole resistance.Methods ERG11 genes of 24 strains Candida albicans isolated from patients were amplified by PCR and bilaterally sequenced.The sequencing result was analyzed by Blast and compared with published sequence (GenBank AY856352) to analyze gene mutations.Then the total RNA was extracted from 24 strains Candida albicans and cDNA was synthesized.The expression of ERG11 gene was detected by fluorescence quantitative RT-PCR.Results 24 silent mutations and 5 missense mutations were identified in 24 strains Candida albicans isolates.Among missense mutations,E174A,T123I and V234F were not found yet in previous studies.Higher expression of ERG11 gene was observed in fluconazoleresistant isolates compared with fluconazole-susceptible isolates [(3.89 ± 2.4) vs.(0.78 ± 0.31)],with statistically significant difference (P ＜ 0.05).Conclusions Drug resistance of Candida albicans to fluconazole may be correlated with the mutations of A114V,E174A,T123I,Y132H,and V234F in ERG11 gene.Besides of this,it is likely that over-expression of ERG11 gene is relevant to fluconazole resistance in Candida albicans.
