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[利用聚合酶链反应特异性引物和DNA探针鉴定白色念珠菌]

[Identification of Candida albicans by specific primers of polymerase chain reaction and DNA probes].

作者信息

Huang C C, Tsai W C, Hseu R S, Wang H H

机构信息

Food Industry Research and Development Institute, Hsinchu, Taiwan, R.O.C.

出版信息

Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi. 1997 Feb;30(1):18-31.

Abstract

Candida albicans is a pathogenic yeast. Two sets of universal primers were used for specific identification of Candida albicans with PCR-amplified ribosomal DNA internal transcribed spacers (ITS). Among the species of Candida, the amplified ITSI and ITSII of DNA fragments were similar in size. The PCR product was purified and labeled with digoxigenin and used as DNA probe in the detection with target DNA of Candida albicans by hybridization. Two sets of specific primers (CA1 and CA2 to amplify ITSI, CA3 and CA4 to amplify ITSII) were designed by alignment of ribosomal ITS sequence of pathogenic Candida albicans with other species to detect C. albicans by PCR. The sensitivity of PCR using the specific primers to detect pure culture of C. albicans was 0.1 ng (about 10(3)-10(4) cells). If the yeast cells were mixed with two other strains, there was a 10-fold decrease in sensitivity (1 ng or 10(4)-10(5) cells) under the same PCR conditions.

摘要

白色念珠菌是一种致病性酵母。使用两组通用引物通过PCR扩增核糖体DNA内部转录间隔区(ITS)对白色念珠菌进行特异性鉴定。在念珠菌属物种中,扩增的DNA片段的ITS1和ITS2大小相似。将PCR产物纯化并用洋地黄毒苷标记,用作DNA探针,通过杂交检测白色念珠菌的靶DNA。通过将致病性白色念珠菌与其他物种的核糖体ITS序列比对,设计了两组特异性引物(CA1和CA2用于扩增ITS1,CA3和CA4用于扩增ITS2),通过PCR检测白色念珠菌。使用特异性引物进行PCR检测白色念珠菌纯培养物的灵敏度为0.1 ng(约10³ - 10⁴个细胞)。如果酵母细胞与另外两种菌株混合,在相同PCR条件下灵敏度会降低10倍(1 ng或10⁴ - 10⁵个细胞)。

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