固相杂交技术检测真菌性角膜病常见致病菌的初步研究.pdfVIP

山东眩药 2006 年第 46 卷第 36 期 回相杂交技术检测真菌性角膜病常见 致病菌的初步研究 张英朗lA ，王丽姬1 赞，李智谛飞到1 声桃l (1 河南省眼科研究所，河南郑州 45003;2 河南省生物工农中心) [摘要] 目的 探讨快速、准确检测真菌性角膜病常见致病盲目的方法。方法 将临床上常见的角膜致病真菌 6属12种经氨基化修饰的特异性寡核昔酸探针固定在隆基化的载破片上，用一对黄光标记通用引物对上述真菌的 DNA 基因片断进行扩增，将带有荧光标i己的扩增产物与载玻片上排列的探针进行杂交，置于荧光显微镜下观察 其且是色情况。结果 12种真商都产生了530~630 坤的 PCR 扩增产物，在相同的条件下对其进行杂交检测，得到 了具有各自特征的杂交后荧光显色图谱，通过荧光倍号可直接对不同商种进行眩分判断。绩论 采用困相杂交技 术能够在3~4 h 完成对临床上常见的12种角膜致病真蔚的蔚种检测。 [关键词〕 固相杂交技术;角膜病;真菌;致病菌;菌种检测 〔中朋分类每] 民772.2 [文献标识码] A [文章编号] 1002精 266X (2006) 36-0017-02 Preliminary study on solid-phase hybridization for detection and identification 01 pathogens spaces in fungal keratitis 1 ZHANG Ying-lang • WANG Liγa.LI Zhiωtao.SHUN Sheng-tao (l Henan Eye Instìtute ,Zhengzhou 450003 ,P. R. China) Abstract: [Objectìve ]丁o develop a technology for the rapid detectìon anò i 也ntìficatìon of common pathog巳ns causing fungal keratitis. [Methods丁 Fixed amino-modified specific oligonucleotide probes to 12 spaces of fungi on a aldehyde slide. These fungal DNA gene fragments were amplified by PCR with one pair of primers labeled with f1 uorescence , and the products were hybridized with probes on slide , tben observed them under fluorescence microscope. [ResultJ 530 - 630 bp PCR amplifìcatìon products developed the visible bands on ag且rose gel electrophoresîs. The fluorescence signal could be found în the aldehyde slide ,which could identify the fungal spaces. [ConclusìonJ Solid-phase hybridization can detect and identjfy pathogens spaces causing fungal keratitis in 3-4 h. Key words: solid-phase hybrìdization; fungal keratitis; fungus; pathogen; identification 菌、白色念珠菌。②主要仪器:PTC町 200 型基因扩增 真菌性角膜病是严重致富01{病之一，快速、敏 仪，