应用M13引物PCR指纹法分析假丝酵母菌氟康唑耐药性

Fluconazole resistance in Candida albicans assayed by PCR fingerprinting with M13 primer

目的探讨M13微卫星引物PCR指纹法用于假丝酵母菌氟康唑耐药性基因分型的可行性。方法采用纸片扩散法分析假丝酵母菌氟康唑耐药性,并进一步对41株假丝酵母菌进行M13引物PCR指纹分析,采用RAPD200软件邻接法(neighbour joining,NJ)聚类分析电泳带型模式。结果41例主要从痰液和阴道分泌物中分离的假丝酵母菌标本中中,氟康唑药物敏感11株(26.8%),依赖8例(19.5%),耐药22例(53.7%),PCR指纹分析至少可检测到12种不同的条带,条带数目从2条到12条不等,电泳带型模式与感染部位和氟康唑药物反应性有关。结论假丝酵母菌氟康唑耐药性菌珠所占比例较大,M13微卫星引物PCR指纹法可用于假丝酵母菌的分子流行病学调查和氟康唑耐药性基因分型。

Objective TounderstandthemolecularandgeneticmechanismunderlyingfluconazoleresistanceinCandida albicans by PCR fingerprinting with M13 primer. Methods Paper disc diffusion method was employed for assay of fluconazoleresistance in 41 clinical isolates of Candida albicans, followed by PCR fingerprinting with M13 primer to study the gel patternswith cluster analysis using neighbor joining (NJ) method performed with RAPD200 software. Results Of the 41 clinicalisolates, 11 strains (26.8%) were fluconazole-sensitive, 8 (19.5%) fluconazole-dependent and 22 (53.7%) flucona-zole-resistance. Two to twelve bands could be observed among these strains, and the gel patterns revealed by cluster analysiswere associated with the reactions of the strains against fluconazole and the location of infection. Conclusion There is highprevalence of fuconazole resistance in clinical Candida albicans isolates, and PCR fingerprinting with M13 primer is con-venient for assay of fuconazole resistance and molecular epidemiological study of Candida albicans.