马内菲青霉菌的鉴定及药敏分析  

Identification of Penicillium Marneffei and Resistance Analysis

作　　者：周俊英[1] 涂建成[1] 郭清莲[1] 

机构地区：[1]武汉大学中南医院检验科,武汉430071

出　　处：《现代检验医学杂志》2011年第4期86-89,共4页Journal of Modern Laboratory Medicine

摘　　要：目的设计马内菲青霉菌特异性引物，探讨马内菲青霉菌病早期诊断的方法及抗生素治疗的药物选择．方法马内菲青霉菌按常规方法放在25℃和37℃进行真茼培养；参照NCCLSM27-A，M38-P对3株马内菲青霉茼的酵母相和茼丝相进行最小抑菌浓度（MIC）测定；PCR的鉴定采用马内菲青霉菌18SrRNA的特征序列，设计内引物Pm1，Pm2和外引物FSF，FSR进行巢式-PCR扩增，并对产物进行产物鉴定。结果马内菲青霉菌为双相性真茼，25℃为青霉相，在沙保弱琼脂平板上产生具有特征性的红色色素，37℃为酵母相，无色素产生；微量稀释法比较马内菲青霉菌在25℃菌丝相和37℃酵母相时的MIC值，结果显示伊曲康唑敏感性最强，其次是酮康唑、5-氟胞嘧啶和氟康唑，最弱的为二性霉素B，3株马内菲青霉菌经巢式-PCR扩增后大约有400bp片段，其他真茼如白色假丝酵母茼、热带假丝酵母茼、由霉菌则没有。结论巢式-PCR可以快速鏊定马内菲青霉菌，48h培养的单个菌落、10h可以鉴定出蛄果，而培养则需4～7d，巢式-PCR对马内菲青霉菌病早期诊断有重要的临床指导意义。医生应结合临床症状及早应用抗真菌药物。Objective To design typical premiers of Penicillium marneffei,to explore the early-phase detection method of Penicillium marneffei infection and to screen the optimal antibiotic treatment. Methods Penicillium marneffei was cul- tured in 25℃ and 37℃ based on theconventional methods,the minimum inhibitory concentration （MIC） of 3 clinical isolates yeast phase and hypha phase was measured by NCCLS M27-A,M38-P methods. Nested PCR identification was＇carried out by designing the outer primers FSF and FSR and inner primers Pml and Pro2 based on the 18s rRNA typical sequence. The PCR products were detected by electrophoresis. Results Penicillium marneffei belonged to double-phase strains, hypha phase in 25℃ with characteristically red pigments in Sabonraud＇s Agar plates,yeast phase in 37℃ ,no red pigments,but with yeast cell morphology. MICs results showed that the drugs sensitive to Penicillium marneffei were Itraconazole,Keto- conanole, 5-Flucylosine, Flueonazol and Amphotercin, respectively, from high concentration to low concentration. Three clinical isolates had 400bp product of nested PCR,but the other fungi such as Candida albicans,Candida tropicalis and AspergiUus had not. Conclusion Penicillium marneffei could be rapidly identified by nested PCR,to isolate single colony spends 48 hours and nested PCR identification took 10 hours,but the culture usually took 4-7 days. Thus the nested PCR technology made senses for the early-phase diseases diagnosis and clinical treatment. The clinical doctor could prescript the antifungi medicine as soon as possible by symptoms and experiment support

关 键 词：马内菲青霉菌 双相真菌 巢式-PCR 药敏试验 

分 类 号：R379[医药卫生—病原生物学] R446.5[医药卫生—基础医学]