吸毒人群口腔念珠菌的DNA多态性及其与耐药性的分析

目的 了解吸毒人群口腔念珠菌的DNA多态性以及与耐药性的关系.方法 选择4种随机引物对75株吸毒人群口腔念珠菌进行DNA多态性分析,并优化试验条件,用聚类分析方法对其进行基因分型.结果 筛选出两条随机引物进行RAPD,念珠菌基因组DNA在100～200 ng/μL,引物浓度在5～10 μmoL/L时,得到的PCR指纹图最丰富和清楚.聚类分析显示,吸毒人群口腔念珠菌同种不同株间存在着基因多态性,不同种间具有一定的基因型特点,某些耐唑类和耐5-氟胞嘧啶药物菌株单独聚为特殊的基因型.结论 RAPD基因分型可作为念珠菌种间鉴定的参考方法.吸毒人群口腔念珠菌中某些菌株的耐药性与其DNA多态性有一定关系,某些特殊PCR指纹图可能与耐唑类药物基因和5-FC基因有一定关系.

analysis on DNA polymorphism and the drug resistance of oral candida species from drug abusers

OBJECTIVE To explore the DNA polymorphism and relationship between drug susceptibility and DNA polymorphism of oral Candida species from drug abusers.METHODS 75 strains of oral candida were selected,and 4 primers were used for analyzing the DNA polymorphism by RAPD and the experiment condition of RAPD technique was optimized.PCR fingerprintings were analyzed for cluster analysis.RESULTS Two pieces of random primer were selected for RAPD-PCR.The fingerprintings were more clearer and more abundant when the genome DNA was 100-200ng/μL and the primer was 5-10μmol/L.It was shown by cluster analysis that there was genetic polymorphism among different stains of same species and there were some certain genotypic characters among different Candida species from drug abusers’ oral cavity.Some resistant strains for azoles and 5FC were clustered into special genotype.CONCLUSION RAPD genotype can be as a referenced method to identify different Candida species.There is a likely relationship between drug resistance of some oral Candida strains from drug abusers and DNA polymorphism.Some distinctive PCR fingerprintings of Candida strains are probably relevant to 5-FC and azoles-resistant genes.