应用实时荧光PCR技术检测烟曲霉的初步研究  

Development of Real-time PCR Method to Detect Aspergillus fumigates

作　　者：刘金华[1,2] 史艳宇[3] 贺丹[1] 刘攀[1] 张宇[1] 徐立波[1] 王丽[1] 

机构地区：[1]吉林大学基础医学院病原生物学教研室,吉林长春130021 [2]吉林出入境检验检疫局,吉林长春130062 [3]吉林省产品质量监督检验院,吉林长春130022

出　　处：《生物技术》2009年第1期34-36,共3页Biotechnology

基　　金：国家自然科学基金项目资助(30571773)

摘　　要：目的:根据烟曲霉Mto1基因特异位点设计并合成探针及引物,建立应用实时荧光PCR检测烟曲霉的方法。方法:通过对多种病原曲霉Mto1基因序列的比对分析,在烟曲霉特异位点设计引物及探针,并对其进行特异性及敏感性进行验证。结果:通过对曲霉属26株不同曲霉菌及其他属的6株不同病原真菌的特异性验证未发现有交叉反应,敏感性实验显示应用该方法可检出1.08×10-6μg/ml的模板DNA。结论:实验建立了应用实时荧光PCR技术检测烟曲霉的方法,该方法具有特异、灵敏、快速等特点并能有效避免普通PCR中的样品间交叉污染问题。A real - time PCR method for the detection of AspergiUusfumigates was established. The primers and TaqMan probes were derived from A. fumigatus - specific sequence of the mitochondrial Mtol gene. 26 Aspergillus isolates and 6 others pathogen fungi strains were used, No cross - amplification was observed. The lowest detection limits of the real - time PCR are proved by the sensitivity testing was 1.08 × 10^-6μg/ml of the A. fum/gates DNA template. Real- time PCR is a time- efficient, specifically, effective and easy - to - use, it is a novel tool for identifying and determining the strain of A. fumigates. This method may be useful for epidemiological studies and surveillance of Aspergillus fumigates in clinical samples.

关 键 词：烟曲霉 实时荧光PCR 检测 

分 类 号：Q93-332[生物学—微生物学]