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当前位置: 首页 > 文章 > 纳米金基因芯片结合通用PCR同时检测多个病原性真菌 生物技术通报 2009,0 (3) 119-122
Position: Home > Articles > Detection Pathogenic Fungi by Nanogold-baesd DNA Microarray Combining with Universal PCR Biotechnology Bulletin 2009,0 (3) 119-122

纳米金基因芯片结合通用PCR同时检测多个病原性真菌

作  者：
鲁卫平;涂植光
单  位：
第三军医大学大坪医院野战外科研究所分子生物学中心;重庆医科大学检验医学系
关键词：
纳米金;基因芯片;聚合酶链反应;真菌
摘  要：
基于致病性真菌rDNA基因分型技术,构建采用通用引物的一次PCR法,并结合纳米金新型基因芯片检测系统实现一次对多个临床常见致病性酵母菌的检测分析。用生物素标记的通用引物扩增各真菌DNA片段并与基因芯片杂交,然后将链酶亲和素标记的纳米金结合到杂交体上,杂交信号通过银染反应被放大形成裸眼可见的显色信息。结果表明,通用引物可扩增临床常见的真菌DNA,选用的各探针特异性强、可靠性好,芯片的最低检测值达50 fmol/L。临床样品检测结果与常规鉴定方法结果一致。该技术检测时间短、操作简单、不需要特殊设备,能部分满足临床检测的通量要求,具有很好的临床应用前景。
译  名：
Detection Pathogenic Fungi by Nanogold-baesd DNA Microarray Combining with Universal PCR
作  者：
Lu Weiping1,2 Tu Zhiguang1(1Faculty of Laboratory Medicine,Chongqing University of Medical Sciences,Chongqing 400016;2Molecular Biology Centerof Daping Hospital of Third Military Medical University,Chongqing 400042)
关键词：
Nano-gold DNA microarray Polymerase chain reaction Medical fungi
摘  要：
A sort of preparation technique that involved in one-time PCR with universal primers and combined with the new detection system of nanogold-baesd gene chip was developed to detect and identify pathogenic yeast.The pathogenic yeast DNA gene fragments were amplified by PCR with universal primers labeled with biotin,and the amplifiers were hybridized with probes on DNA microarray.Then the streptavidin-gold was integrated into the hybrid complex.Hybridization signal was amplified by the interaction between nanogold and silver stain reagent to form spot which could be detected with naked eyes.The detection results of clinical sample were consistent with that of obtained by traditional microbiological culture and biochemical techniques.It proved that this method is a simple,fast,sensitive technology and no special equipment should be needed.It can meet the flux demand of clinical detection and has good prospect of clinical application.

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