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白色念珠菌DNA探针的研制

Development of DNA probes for Candida albicans.

作者信息

Cheung L L, Hudson J B

机构信息

Division of Medical Microbiology, University of British Columbia, Vancouver, Canada.

出版信息

Diagn Microbiol Infect Dis. 1988 Jul;10(3):171-9. doi: 10.1016/0732-8893(88)90037-5.

Abstract

An attempt was made to produce DNA probes that could be used as a rapid and efficient means of detecting candidiasis (invasive Candida infection) in immunocompromised patients. Whole DNA from Candida albicans was digested with restriction endonuclease, and the resulting fragments were randomly cloned into a plasmid vector. Several recombinant plasmids were evaluated for cross-hybridization to various other Candida species, other fungal DNAs, and to nonfungal DNAs. Cross reactions were observed between the probes and different yeasts, but none with unrelated DNAs. Some recombinants were genus-specific, and two of these were applied to the analysis of C. albicans growth curves. It became evident that, although both 32P- and biotin-labelled probes could be made quite sensitive, a possible limitation in their diagnostic potential was the poor liberation of Candida DNA from cells. Thus, better methods of treatment of clinical specimens will be required before such probes will be useful in routine diagnosis.

摘要

研究人员试图制备DNA探针,用作检测免疫功能低下患者念珠菌病(侵袭性念珠菌感染)的快速有效方法。白色念珠菌的全基因组DNA用限制性内切酶消化,所得片段随机克隆到质粒载体中。评估了几种重组质粒与其他多种念珠菌属物种、其他真菌DNA以及非真菌DNA的交叉杂交情况。在探针与不同酵母之间观察到交叉反应,但与不相关的DNA无交叉反应。一些重组体具有属特异性,其中两个被用于分析白色念珠菌的生长曲线。显而易见,尽管32P标记和生物素标记的探针都可以具有相当高的灵敏度,但其诊断潜力的一个可能限制是念珠菌DNA从细胞中释放不佳。因此,在这类探针可用于常规诊断之前,需要更好的临床标本处理方法。

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