中文摘要：

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。

英文摘要：

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.