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用任意引物聚合酶链反应鉴别创伤弧菌菌株

Differentiation of Vibrio vulnificus strains by an arbitrarily primed polymerase chain reaction.

作者信息

Wu J J, Hor L I, Shiau S L

机构信息

Department of Medical Technology, National Cheng Kung University Medical College, Taian, Taiwan, R.O.C.

出版信息

Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi. 1995 Feb;28(1):70-8.

PMID:9774986
Abstract

A synthetic 17 mer oligonucleotide (5'-GTTGGGTAACGCCAGGG-3') was used as a primer for the arbitrarily primed polymerase chain reaction (AP-PCR) to differentiate various strains of Vibrio vulnificus. A total of 37 genomic DNAs that were extracted from the clinical and environmental strains were successfully differentiated. Among them, 32 profiles of the 37 strains were characterized. None of the environmental and clinical strains had the same amplification profile, suggesting the highly heterogeneous population existed in the strains of V. vulnificus. The size of the amplified sequences ranged from 0.3 to 2.0 Kb and the DNAs were separated to 12 to 20 bands by the 1.2% agarose gel. The clinical isolates from two independent episodes of V. vulnificus infections in a patient were shown to have the same profile, indicating that the second episode was due to recurrence rather than reinfection. The profiles of amplification were reproducible with different preparations of genomic DNA. Arbitrarily primed polymerase chain reaction can therefore be a useful tool for epidemiological study of V. vulnificus infection.

摘要

一条合成的17聚体寡核苷酸(5'-GTTGGGTAACGCCAGGG-3')被用作任意引物聚合酶链反应(AP-PCR)的引物,以区分不同的创伤弧菌菌株。从临床和环境菌株中提取的总共37个基因组DNA成功地被区分开来。其中,37个菌株中的32个图谱得到了表征。环境菌株和临床菌株均没有相同的扩增图谱,这表明创伤弧菌菌株中存在高度异质性群体。扩增序列的大小范围为0.3至2.0 kb,DNA通过1.2%琼脂糖凝胶分离为12至20条带。一名患者两次独立的创伤弧菌感染临床分离株显示具有相同的图谱,表明第二次感染是由于复发而非再感染。不同基因组DNA制剂的扩增图谱具有可重复性。因此,任意引物聚合酶链反应可成为创伤弧菌感染流行病学研究的有用工具。

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