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目的分离妇科阴道炎患者感染的白念珠菌,检测其耐药性及耐药株基因突变情况,以指导临床合理用药。方法取临床分离的白念珠菌耐药株和敏感株,经传代后再次进行药敏试验,以确定耐药情况。然后提取基因组DNA,进行电泳检测;采用PCR扩增ERG11基因并测序。结果药敏试验显示氟康唑对白念株菌GZ16传代后的MIC值为64μg/ml,判定GZ16为耐药株;SC5314的MIC值为2μg/ml,判定为敏感株;基因组DNA电泳,可见白念株菌GZ16、SC5314两菌株目的条带位于相同位置;对ERG11基因测序,在GZ16白念珠菌有G487T和T916C两个突变位点,SC5314白念珠菌有All67G、A1587G及T462A、T495A、A504G、A530C、C558T、C805T等8个基因位点的突变,并且还存在氨基酸的错义突变,包括:F105L(T462A)、D116E(T495A)、K128T(A530C)。结论分离自妇科阴道炎患者的白念珠菌部分对氟康唑耐药并发生ERG11基因G487T和T916C位点突变,该突变可能与白念珠菌对氟康唑耐药有关。
Objective To isolate Candida albicans infected with gynecological vaginitis and test its drug resistance and gene mutation of drug-resistant strains to guide clinical rational drug use. Methods Candida albicans isolates and sensitive strains were isolated from clinical isolates and re-tested for susceptibility to determine drug resistance. Then genomic DNA was extracted for electrophoresis detection; ERG11 gene was amplified by PCR and sequenced. Results The susceptibility test showed that the MIC value of fluconazole against GZ16 was 64μg / ml, GZ16 was the resistant strain, and the MIC value of SC5314 was 2μg / ml, which was judged as sensitive strain. Genomic DNA electrophoresis showed that white Candida strains GZ16, SC5314 two strains located in the same location; sequencing of ERG11, Candida albicans GZ16 G487T and T916C two mutation sites, SC5314 Candida albicans have All67G, A1587G and T462A, T495A, A504G, A530C, C558T, C805T and other 8 loci mutations, and there are also amino acid missense mutations, including: F105L (T462A), D116E (T495A), K128T (A530C). Conclusion Candida albicans isolated from patients with gynecologic vaginitis is resistant to fluconazole and mutations in G487T and T916C sites of ERG11 gene may occur. This mutation may be related to the resistance of Candida albicans to fluconazole.