聚合酶链反应特异性引物与DNA探针鉴定白色念珠菌[j]。

C C Huang, W C Tsai, R S Hseu, H H Wang
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引用次数: 0

摘要

白色念珠菌是一种致病酵母菌。采用两组通用引物对白色念珠菌进行pcr扩增核糖体DNA内转录间隔物(ITS)特异性鉴定。在念珠菌种间,DNA片段的ITSI扩增值与ITSI大小相近。PCR产物纯化后用地高辛标记,作为DNA探针与白色念珠菌靶DNA杂交检测。通过将致病性白色念珠菌核糖体ITS序列与其他菌种比对,设计了两组特异性引物(CA1和CA2扩增ITSI, CA3和CA4扩增ITSII),采用PCR检测白色念珠菌。特异引物PCR检测纯培养白色念珠菌的灵敏度为0.1 ng(约10(3)-10(4)个细胞)。如果酵母细胞与其他两种菌株混合,在相同的PCR条件下,灵敏度降低了10倍(1 ng或10(4)-10(5)个细胞)。
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[Identification of Candida albicans by specific primers of polymerase chain reaction and DNA probes].

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.

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