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与无胸腺裸鼠角化过度性皮炎相关的棒状杆菌16S rRNA基因序列分析及基于PCR的检测方法的建立。

Analysis of the 16S rRNA gene sequence of the coryneform bacterium associated with hyperkeratotic dermatitis of athymic nude mice and development of a PCR-based detection assay.

作者信息

Duga S, Gobbi A, Asselta R, Crippa L, Tenchini M L, Simonic T, Scanziani E

机构信息

Istituto di Fisiologia Veterinaria e Biochimica, Università di Milano, Italy.

出版信息

Mol Cell Probes. 1998 Aug;12(4):191-9. doi: 10.1006/mcpr.1998.0168.

Abstract

By 16S rDNA sequencing the authors have characterized the coryneform bacteria associated with hyperkeratotic dermatitis (HD) of athymic nude mice isolated from six different outbreaks of the disease in Northern Italy. This analysis has allowed the authors to confirm the classification of the bacteria as Corynebacterium bovis and to develop a 16S rDNA-based polymerase chain reaction (PCR) detection assay. The test was performed directly on the DNA extracted from epidermal swabs. The PCR primers were chosen to match the 16S rDNA sequence fragments which differ most from the other Corynebacterium spp. The test was shown to be both sensitive and specific for C. bovis. Detection of as few as three viable bacterial cells was possible with the use of an oligonucleotide probe in a liquid hybridization assay.

摘要

通过16S rDNA测序,作者对从意大利北部六次不同疾病暴发中分离出的无胸腺裸鼠的角化过度性皮炎(HD)相关棒状杆菌进行了特征分析。该分析使作者能够确认这些细菌为牛棒状杆菌,并开发了一种基于16S rDNA的聚合酶链反应(PCR)检测方法。该检测直接对从表皮拭子中提取的DNA进行。选择PCR引物以匹配与其他棒状杆菌属差异最大的16S rDNA序列片段。结果表明该检测方法对牛棒状杆菌具有敏感性和特异性。在液体杂交试验中使用寡核苷酸探针,能够检测到低至三个活细菌细胞。

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