摘要
目的 针对血液病真菌感染日益严重趋势 ,建立念珠菌基因扩增技术并应用于临床。方法 本研究应用Lyticase破真菌壁提取DNA ,采用二对引物分别对白色念珠菌EO3 12 5bpDNA和念珠菌细胞色素P4 5 0L1A12 4 3bpDNA进行基因扩增 ;并分别对白色念珠菌标准株、临床样本株、其他念珠菌和不同细菌菌株进行基因扩增特异性比较 ,同时应用该技术检测血液病患者痰液和血液中念珠菌。结果 基因扩增显示EO3 12 5bpDNA只有标准株和临床样本白色念珠菌阳性 ,细胞色素P4 5 0L1A12 4 3bpDNA具有念珠菌种的特异性 ,临床血液和痰液标本检测阳性率显著提高 ,与临床病情一致。结论 血液病念珠菌感染基因扩增技术敏感性、特异性显著增强 ,而且耗时显著缩短 。
OBJECTIVE DNA amplification was established and used for investigation of clinical Candida infections with hematological diseases. METHODS Application of the polymerase chain reaction to the diagnosis of candidosis by amplification of EO 3 125 bp (from mitochondrial DNA) and cytochrome P 450 L1A1 (lanosterol-14а-demethylase) gene fragment in cases with hematological diseases. RESULTS The 125 bp DNA from EO 3 gene of Candida albicans was only amplified from standard strain of C. albicans and clinical samples from C. albicans infection cases with hematological diseases, meanwhile, cytochrome P 450 L1A1 gene fragment could be amplified from C. albicans, C. tropicalis and C. parapsilosis including standard strains and clinical samples. CONCLUSIONS The 125 bp DNA from EO 3 gene is specific and sensitive to C. albicans and cytochrome P 450 L1A1 gene fragment to C. albicans. Genetic diagnosis of C. albicans would be useful and suitable for the rapid identification of clinical candidosis in cases with hematological diseases.
出处
《中华医院感染学杂志》
CAS
CSCD
2003年第10期904-908,共5页
Chinese Journal of Nosocomiology
基金
福建省自然科学基金资助 (课题编号 94 0 1 批准号 94 0 1)
福建省医药卫生基金资助 (课题编号970 6 批准号 96 0 31)
关键词
念珠菌属
基因扩增
血液病
Candida
DNA amplification
Hematological diseases
