中华皮肤科杂志 ›› 1993, Vol. 26 ›› Issue (2): 96-98.

• 论著 • 上一篇    下一篇

光生物素标记念珠菌基因探针的研究

李春阳1, 吴绍熙2, 郭宁如2, 李银太3, 林万明3   

  1. 1. 山东医科大学附属医院皮肤科, 济南, 250012;
    2. 中国医学科学院皮肤病研究所;
    3. 军事医学科学院徽生物流行病研究所
  • 收稿日期:1991-02-18 修回日期:1992-07-20 出版日期:1993-04-15 发布日期:1993-04-15

Study on candida albicans DNA probe latelled with photobiotin

LI Chun-Yang1, WU Shao-Xi2   

  1. The Affiliated Hospital of Shandong Medical University, Ji'nan 250012
  • Received:1991-02-18 Revised:1992-07-20 Online:1993-04-15 Published:1993-04-15

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摘要： 采用6-氨基己酸光生物素(EACA-Photobiotion)和生物素交联补骨脂素(Biotin-psoralea)标记白念珠菌标准株Cli DNA 2.5kb片段为探针,与各种念珠菌,隐球菌和细菌进行核酸杂交,并与32P标记探针作了比较.经亲和素-碱性磷酸酶系统(Avidin-alkaline phosphatase,A-AP)显色,6-氨基己酸光生物素的标记活性为1pg,其标记探针可检测出100pg以下的靶DNA,与全细胞DNA杂交,检测敏感性为125pg,较生物素交联补骨脂素的标记活性和检测敏感度约高10倍.两种光生物素标记探针与白念珠菌杂交均显示强阳性信号,与5株近平滑念珠菌和1株热带念珠菌有微弱杂交信号,与其它几种常见致病性念珠菌、隐球菌和细菌不杂交.结果表明6-氨基己酸光生物素标记白念珠菌CliDNA2.5kb探针对白念珠菌的检测敏感度和特异性与同位素标记探针基本一致,并且具有操作简便,快速,安全及探针使用期长、价廉等优点.

关键词: 光生物素, 白念珠菌, 探针

Abstract: 2.5kb DNA fragment from a standard strain of Candida albicans (C li) was labeled with E-AGA-Photobiotin and Biotin-psoralen, respectively, and used as probe to hybridize with DNA from various Candida species, Cry-ptococcus and bacteria. Colorimetric sensitivity of E-ACA-Photobiotin labeled probe with Avidin-Alkaline phosphatase (A-AP) is 1 pg. Detection of target DNA by labeled probe is less than 100pg, and that of whole cellular DNA is 125pg, being ten times higher than that of the probe labeled with biotin-psoralen. 7 strains of C. albicans DNA hybridized with the above two photohiotin labelled probes gave strong positive results,and C. parapsiiasis showed weak hybridization signal. DNA from other Candida species, Gryptococcus and bacteria did not hybridize with the probes. These results showed that the sensitivity and specificity of C. albicans Clin 2.5kb probe labeled with E-ACA-Photobiotin are in conformity with that of radioactive labelling one. Moreover, with the advantages of simplicity, safety, stability and rapidity, photobiotin labelling method can replace the isotope labelling method.

Key words: Candida albicans, Probe, Photahiotin