探讨白念珠菌基因多态性与耐药性关系
2014-01-29 02:35
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【摘要】 【目的】 探讨耐氟康唑白念珠菌基因多态性与其耐药性是
【摘要】 【目的】 探讨耐氟康唑白念珠菌基因多态性与其耐药性是否相关。【方法】 收集48株敏感白念珠菌和10株耐氟康唑白念珠菌，其中包括2株体外诱导的耐氟康唑白念珠菌。选择1个随机引物（引物1251），采用任意引物PCR方法基因分型，将电泳图谱扫描入计算机后采用Labwork4.0软件转化为数值图表,利用SPSS 11.5进行聚类分析。 【结果】 引物1251的PCR指纹图带型稳定，多态性丰富，可以作为分型引物。聚类分析结果提示8株临床耐药菌的PCR指纹图相似系数为71.7%，而其中7株的耐药菌相似系数高达86.7%，远高于58株菌的平均相似系数(47.3%)。2株体外诱导的耐药菌诱导前后基因型未发生变化。【结论】 基因分型的结果未提示耐氟康唑白念珠菌存在一定的特殊电泳条带，但提示了特定的PCR指纹图可能与耐药性有一定相关性。
【关键词】 白念珠菌； 基因型； 氟康唑； 耐药性，任意引物PCR
Abstract： 【Objectives】 To investigate the possible relationship of C. albicans gene polymorphisms and its fluconazole-resistance. 【Method】 Forty-eight fluconazole-susceptible stains and 10 fluconazole-resistant stains of C. albicans were analyzed, with two resistant stains induced in vitro. All of them were genotyped by arbitrary primed polymerase chain reaction (AP-PCR) fingerprinting employing one interrepeat primes (1251 primers). The image was captured using computer-assisted system with Labwork 4.0 software to analyze the gel patterns of all isolates. The data were processed by SPSS 11.5 statistical software. 【Results】 Primer 1251 was suitable for fingerprinting analysis, and was found to generate the reproducible fingerprinting profiles, yielding well-resolved banding patterns. The AP-PCR profiles of 8 resistant stains isolated clinically were highly similar according to similarity coefficient (71.7%). Furthermore, the similarity coefficient of 7 resistant isolates (86.6%) was significantly higher than that of total 58 stains (47.3%). For two induced isolates, their genotypes did not change much after resistance-inducement in vitro. 【Conclusions】 No specific DNA profiles of C.albicans indicating fluconazole-resistance were identified. However, our results demonstrated a strong genetic correlation between fluconazole-resistant patterns and its AP-PCR profiles in C.albicans.
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Key words： C.albicans; genotype; fluconazole; drug resistance; AP-PCR
近年来随着免疫抑制剂、抗肿瘤药物、广谱抗生素的应用及艾滋病流行，临床上因白念珠菌感染所致的深部真菌病逐年增多，而过多、广泛的使用抗真菌药物所产生的选择压力，使念珠菌的耐药现象有明显增多的趋势。白念珠菌的耐药表型与基因型的关系目前仍不清楚，寻找一种恰当的基因分型方法，探讨其与耐药表型的关系，对监测耐药株的分子流行病学，以及深化耐药分子机制的研究有重要的临床意义[1]。我们应用任意引物PCR（Arbitary primed -PCR）的方法对临床分离的白念珠菌进行了基因分型，研究白念珠菌的耐药性与基因多态性的关系。
1 材料与方法
1.1 受试菌株
8株耐氟康唑白念珠菌（MIC
