| 用DNA microarray快速检测淋球菌耐喹诺酮类药物基因突变 |
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| 引用本文： | 周文明,赵建龙,杨森,曹慧敏,李伟,沈玉君,张书梅,张学军.用DNA microarray快速检测淋球菌耐喹诺酮类药物基因突变[J].疾病控制杂志,2004,8(2):106-108. |
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| 作者姓名： | 周文明 赵建龙 杨森 曹慧敏 李伟 沈玉君 张书梅 张学军 |
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| 作者单位： | 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,安徽,合肥,230022
2. 中国科学院上海微系统与信息技术研究所,上海,200050 |
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| 基金项目： | 中国科学院生物科学与生物技术研究特别支持经费课题 (STZ- 0 0 - 0 3) |
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| 摘 要： | 研究DNA microarray的制备及其检测淋球菌耐喹诺酮类药物基因突变的准确性。方法根据淋球菌药敏及测序结果分别对淋球菌gyrA和parC基因的序列设计特异引物和探针并制作DNA microarray。对淋球菌临床拭子进行PcR扩增并荧光标记包含gyrA和parC基因的目的DNA片段，与芯片杂交，同时以测序法进行双盲淋球菌耐喹诺酮类药物基因突变的检测。结果87份泌尿生殖道试子全部可用DNA microarray检测出来，芯片检测结果与药敏结果符合率为100％，与测序结果符合率为97．7％。结论用DNA microarray来检测淋球菌gyrA和parC基因突变快速、特异性高和灵敏度高，可以应用于临床耐药性检测。
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| 关 键 词： | DNA microarray 检测 淋球菌 喹诺酮类药物 基因突变 耐药基因 |
| 文章编号： | 1008-6013(2004)02-0106-03 |
| 修稿时间： | 2003年10月15 |
Rapid detection of the mutations of resistance-fluoroquinolone neisseria gonorrhoeae gene by a DNA microarray |
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| ZHOU Wen-ming ,ZHAO Jian-long ,YANG Sen ,CAO Hui-min ,LI Wei ,SHEN Yu-jun ,ZHANG Shu-mei ,ZHANG Xue-jun.Rapid detection of the mutations of resistance-fluoroquinolone neisseria gonorrhoeae gene by a DNA microarray[J].Chinese Journal of Disease Control and Prevention,2004,8(2):106-108. |
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| Authors: | ZHOU Wen-ming ZHAO Jian-long YANG Sen CAO Hui-min LI Wei SHEN Yu-jun ZHANG Shu-mei ZHANG Xue-jun |
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| Affiliation: | ZHOU Wen-ming 1,ZHAO Jian-long 2,YANG Sen 1,CAO Hui-min 2,LI Wei 1,SHEN Yu-jun 1,ZHANG Shu-mei 1,ZHANG Xue-jun 1. Department of Dermatology,the First Affiliated Hospital of Anhui Medical University,Hefei 230022,China, 2.Shanghai Institute of Microsystem and Information Technology,Chinese Academy of Sciences,Shanghai200050,China |
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| Abstract: | Objective To study the preparation of DNA microarray and its accuracy in detection of the mutations of assistance-fluoroquinolone neisseria gonorrhoeae genes. Methods The primers and probes were prepared according to the sequence of neisseria gonorrhoeae gyrA and parC genes. The DNA fragment of genital swabs which contained gyrA and parC genes were labeled with cy5 fluorescence and amplified by PCR technique. Then they were hybridized with DNA microarray. DNA sequencing data of samples were detected by double blind method. Results 87 strains genital swab sample were detected using DNA microarray. The results were consistant with drug sensitivity test. The double-blind test results showed a coincidence rate of 97.7% between sequencing data and DNA microarray. Conclusions It showed higher speciality and sensitivity using DNA microarray to rapidly detect the mutations of resistance-fluoroquinolone neisseria gonorrhoeae genes. This method could be used for clinical detection of Fluoroquinolone resistance strains. |
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| Keywords: | neisseria gonorrhoeae quinolones genes mutation drug-resistance |
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