中华皮肤科杂志 ›› 2005, Vol. 38 ›› Issue (12): 735-737.

• 论著 • 上一篇    下一篇

检测泌尿生殖道感染常见性病病原体基因芯片的构建

吴黎明1, 孙建方1, 蒋明军1, 刘全俊2, 陈岳明3, 张红河3   

  1. 1. 中国医学科学院、中国协和医科大学皮肤病研究所 南京 210042;
    2. 东南大学吴健雄实验室;
    3. 杭州市第一人民医院
  • 收稿日期:2005-02-01 发布日期:2005-12-15
  • 通讯作者: 孙建方,E-mail:sunjianf@jlonline.com E-mail:sunjianf@jlonline.com

Development of a Gene Chip for the Detection of Common Pathogens Causing Urogenital Sexually Transmitted Infections

WU Li-ming1, SUN Jian-fang1, JIANG Ming-jun1, Liu Quan-jun2, CHEN Yue-ming3, ZHANG Hong-he3   

  1. Institute of Dermatology, Chinese Academy of Medical Sciences, Peking Union Medical College, Nanjing 210042, China
  • Received:2005-02-01 Published:2005-12-15

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摘要： 目的 建立检测泌尿生殖道性传播感染常见病原体的基因芯片技术。方法 首先将9种目的病原体分为病毒、细菌及低级真核生物三类,分别设计1对荧光标记的通用引物,利用多重PCR技术使3对引物置于同一反应体系进行扩增,再将特异性探针点样于玻片上制备成基因芯片,与扩增产物进行杂交、扫描并分析结果。结果 单重PCR和多重PCR扩增产物的琼脂糖凝胶电泳均可见相应的阳性条带。扩增产物与基因芯片杂交后扫描,各病原体均可见荧光信号,与所对应的特异性探针位置相吻合。结论 本研究初步构建的基因芯片技术具有特异、灵敏、快速及能一次性同时检测多种病原体的特点。

关键词: 寡核苷酸序列分析, 性传播疾病

Abstract: Objective To develop a gene chip for the detection of common pathogens causing urogenital sexually transmitted infections. Methods The target pathogens were divided into three groups: viruses, bacteria, and lower eukaryotes. Three pairs of universal primers were designed and applied to amplify the target genes of these different pathogens in one PCR reaction system. The gene chips were then prepared via immobilization of the specific probes onto specially treated glass slides. Finally, the labeled amplicons were hybridized with the gene chips, scanned and analyzed using computer software. Results Amplicons were detected by agarose gel electrophoresis. The fluorescence signals for specific pathogens could be recognized in the gene chips, and were identical with the positions of the specific probes. Conclusions Gene chip is a specific, sensitive and rapid method for simultaneous detection of multiple sexually transmitted infections.

Key words: Oligonucleotide array sequence analysis, Sexually transmitted diseases