固相杂交技术检测真菌性角膜病常见致病菌的初步研究  

Preliminary study on solid-phase hybridization for detection and identification of pathogens spaces in fungal keratitis

作　　者：张英朗[1] 王丽娅[1] 李智涛 孙声桃[1] 

机构地区：[1]河南省眼科研究所 [2]河南省生物工程中心

出　　处：《山东医药》2006年第36期17-18,共2页Shandong Medical Journal

基　　金：河南省杰出人才创新基金项目(0321002000);河南省科技攻关计划项目(0623033000)

摘　　要：目的探讨快速、准确检测真菌性角膜病常见致病菌的方法。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苷酸探针固定在醛基化的载玻片上。用一对荧光标记通用引物对上述真菌的DNA基因片断进行扩增，将带有荧光标记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显色情况。结果12种真菌都产生了530～630bp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，通过荧光信号可直接对不同菌种进行区分判断。结论采用固相杂交技术能够在3～4h完成对临床上常见的12种角膜致病真菌的菌种检测。[Objective] To develop a technology for the rapid detection and identification of common pathogens causing fungal keratitis. [Methods] 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 fluorescence, and the products were hybridized with probes on slide, then observed them under fluorescence microscope. [Result] 530-630 bp PCR amplification products developed the visible bands on agarose gel eleetrophoresis. The fluorescence signal could be found in the aldehyde slide,which could identify the fungal spaces. [Conclusion] Solid-phase hybridization can detect and identify pathogens spaces causing fungal keratitis in 3-4 h.

关 键 词：固相杂交技术 角膜病 真菌 致病菌 菌种检测 

分 类 号：R772.2[医药卫生—眼科]