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目的探讨白色念珠菌upc2基因C末端单核苷酸多态性以及其对唑类药物耐药的影响。方法对34株白色念珠菌临床株,纸片法测定5种药物敏感性,DNA测序法检测upc2基因单核苷酸多态性,RT-PCR法测定cdr1基因m RNA转录水平。比较耐药株和敏感株upc2单核苷酸多态性以及cdr1基因m RNA水平的差异。结果 14株白色念珠菌氟康唑耐药株(3株多重耐药,6株双重耐药,5株单一耐药),20株敏感株对5种药物均敏感。5株耐药白色念珠菌upc2基因存在G1927A型多态性,20株敏感株upc2基因均为G1927野生型,G1927A型upc2基因检出率35.7%,显著高于敏感株(χ2=8.37,P<0.01)。G1927A型多重耐药株的cdr1基因m RNA水平高于G1927A型双重耐药株,并且二者均高于敏感株。结论 upc2基因G1927A单核苷酸多态性通过上调cdr1基因m RNA转录而引起白色念珠菌对唑类药物产生多重耐药或双重耐药表型。
Objective To investigate the C-terminal single nucleotide polymorphisms of the upc2 gene of Candida albicans and its effects on the resistance of azole drugs. Methods 34 strains of Candida albicans clinical isolates were tested, and five drug sensitivity tests were performed by paper method. The single nucleotide polymorphism of upc2 gene was detected by DNA sequencing and the m RNA transcription level of cdr1 gene was determined by RT-PCR. The differences of single nucleotide polymorphisms of upc2 and mrRNA of cdr1 gene between resistant and susceptible strains were compared. Results Fourteen Candida albicans fluconazole-resistant strains (3 multi-drug resistant, 6 double-drug resistant and 5 single drug-resistant) and 20 sensitive strains were sensitive to 5 drugs. The genotypes of upc2 of Candida albicans were G1927A polymorphism. The upc2 genes of 20 strains were all G1927 wild type. The detection rate of G1927A upc2 gene was 35.7%, which was significantly higher than that of the susceptible strain (χ2 = 8.37, P < 0.01). The mrRNA level of cdr1 gene of G1927A multidrug resistant strains was higher than that of G1927A double resistant strains, and both were higher than the sensitive strains. Conclusion The Gc1927A single nucleotide polymorphism of upc2 gene leads to multidrug resistance or double drug resistance phenotypes of Candida albicans to azoles by up-regulating the m RNA transcription of cdr1 gene.