中华皮肤科杂志 ›› 2004, Vol. 37 ›› Issue (5): 259-261.

• 论著 • 上一篇    下一篇

DNA芯片快速检测淋球菌gyrA基因突变

周文明1, 张学军1, 曹慧敏2, 杨森1, 孙良丹1, 赵建龙2   

  1. 1. 安徽医科大学皮肤性病研究所、第一附属医院皮肤性病科, 合肥230022;
    2. 中国科学院上海微系统与信息技术研究所
  • 收稿日期:2003-05-20 出版日期:2004-05-15 发布日期:2004-05-15

Rapid Detection of Mutations of Neisseria gonorrhoeae gyrA Gene Using DNA Chip

ZHOU Wen-ming1, ZHANG Xue-jun1, CAO Hui-min2, YANG Sen1, SUN Liang-dan1, ZHAO Jian-long2   

  1. Department of dermatology, The First Affiliated Hospital of Anhui Medical University, Hefei 230022, China
  • Received:2003-05-20 Online:2004-05-15 Published:2004-05-15

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摘要： 目的 研制一种新型DNA芯片,用于快速检测淋球菌gyrA基因突变。方法 根据淋球菌gyrA基因的序列信息设计探针并制作DNA芯片,PCR扩增并荧光标记包含gyrA基因突变的目的DNA片段,与芯片杂交,同时以测序法为对照。结果 50份泌尿生殖道拭子全部可用DNA芯片检测出来,芯片检测结果与药敏结果符合率为100%,与测序结果二者符合率为98%。结论 DNA芯片检测淋球菌gyrA基因突变具有快速、高特异性和高灵敏度,可以应用于临床耐药性检测。

关键词: 寡核苷酸序列分析, DNA促旋酶, 奈瑟球菌,淋病, 喹诺酮类, 点突变

Abstract: Objective To develop a new method-DNA chip to be used for rapid detection of mutations of Neisseria gonorrhoeae gyrA gene. Methods Probes were designed according to the sequence of Neisseria gonorrhoeae gyrA genes, and DNA chip was fabricated accordingly. DNA fragment which contains gyrA gene mutation was amplified using PCR technique, labeled with Cy5 fluorescence, and then hybridized with DNA chip. Results of DNA sequencing were used as the control. Results All of the 50 urogenital swab specimens were detected using DNA chip. The consistency between the DNA chip and drug sensitivity test, and between the DNA chip and DNA sequencing were 100% and 98%, respectively. Conclusions DNA chip is a rapid technique with high sensitivity and specificity for the detection of mutations of Neisseria gonorrhoeae gyrA gene. This method can be used for the detection of drug resistance in clinical practice.

Key words: Neisseria Gonorrhoeae, Quinolones, Point mutation, DNA gyrase, Oligonucleotide array sequence analysis