中华皮肤科杂志 ›› 2011, Vol. 44 ›› Issue (12): 861-864.

• 论著 • 上一篇    下一篇

白念珠菌芽管特异性抗原快速检测方法的建立和初步评价

赵桂芝1,韩慧霞2,宋秋荷3,钟白玉4,陆洪光5,王鲁6   

  1. 1. 重庆市第一人民医院
    2. 贵阳医学院皮肤病与性病学专业与重庆市第一人民医院联合招生2008级硕士研究生
    3. 江西九江学院临床医学院 皮肤科
    4. 重庆市第三军医大学西南医院皮肤科
    5. 贵阳医学院附属医院皮肤科
    6. 重庆市第一人民医院皮肤科
  • 收稿日期:2010-12-28 修回日期:2011-08-25 出版日期:2011-12-15 发布日期:2011-12-02
  • 通讯作者: 王鲁 E-mail:wlwwh2003@yahoo.com.cn

Establishment and preliminary evaluation of a rapid detection method for germ tube-specific antigens of Candida albicans

guizhi ZHAO1,Hui-Xia HAN1,Qiu he Song1, 1,   

  • Received:2010-12-28 Revised:2011-08-25 Online:2011-12-15 Published:2011-12-02

摘要:

目的 建立白念珠菌芽管特异性抗原快速检测系统,探讨其特异性和敏感性。方法 以单克隆抗体McAb03.2C1-C2作为一抗,辣根过氧化物酶标记McAb03.2C1-C2为二抗,建立双抗体夹心ELISA实验方法并检测6只动物模型和5例系统性白念珠菌感染患者的血清标本。结果 筛选出McAb03.2C1-C2最佳稀释度和检测包被抗原浓度分别为1 ∶ 4000和1.25 ~ 40 μg/ml;双抗体夹心ELISA检测动物模型的特异性和敏感性分别为95%、92%,其对阳性临床标本的检测结果与常规鉴定方法一致。结论 初步建立白念珠菌芽管特异性抗原的双抗体ELISA快速检测系统,动物模型实验证实该系统有较好的特异性和敏感性。

关键词: 敏感性与特异性

Abstract:

Objective To establish a double-antibody sandwich enzyme linked immunosorbent assay(DAS-ELISA) for the rapid detection of germ tube-specific antigens of Candida albicans, and to evaluate its specificity and sensitivity. Methods A DAS-ELISA was established with the monoclonal antibody McAb03.2C1-C2 as the primary antibody, and horseradish peroxidase (HRP) labeled McAb03.2C1-C2 as the secondary antibody. The established assay was used to detect germ tube-specific antigens of Candida albicans in sera from 5 patients with systemic Candida albicans infection and from 6 rabbit models at 12, 24, 48, 72 hours, on week 1, 2 after infection with Candida albicans. Results A good liner relationship was observed between the absorbance value at 495 nm and antigen concentrations when the titer of McAb03.2C1-C2 was 1 ∶ 4000 and the concentration of coated antigen varied from 1.25 to 40 μg/ml. The specificity and sensitivity of the DAS-ELISA were 95% and 92% respectively in the detection of germ tube-specific antigens in the rabbit models. The results of detection with DAS-ELISA in serum specimens from the patients were consistent with those with the routine method. Conclusions A DAS-ELISA is primarily established for the rapid detection of germ tube-specific antigens of Candida albicans, and has shown a satisfactory sensitivity and specificity in the animal model experiment.