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利用肠杆菌重复基因间共有序列(ERIC)基序-聚合酶链反应/毛细管电泳对单核细胞增生李斯特菌进行DNA指纹分析。

DNA fingerprinting of Listeria monocytogenes using enterobacterial repetitive intergenic consensus (ERIC) motifs-polymerase chain reaction/capillary electrophoresis.

作者信息

Sciacchitano C J

机构信息

US Food and Drug Administration, Northeast Regional Laboratory, Brooklyn, NY 11232, USA.

出版信息

Electrophoresis. 1998 Jan;19(1):66-70. doi: 10.1002/elps.1150190112.

Abstract

The molecular technique, enterobacterial repetitive intergenic consensus (ERIC)-polymerase chain reaction (PCR) produces genomic DNA fingerprint that discriminate bacterial species and strains. This technique was applied to the characterization of Listeria monocytogenes, an important food-borne pathogen implicated in numerous cases of listeriosis. The ERIC-PCR resulted in distinct DNA fingerprinting patterns of all L. monocytogene serotypes and Listeria species. Analysis of the genomic DNA fingerprints was accomplished using capillary electrophoresis (CE), an alternative technique to the conventional agarose gel method. The optimization of CE conditions (electrokinetic injection, applied voltage) resulted in the resolution of amplified DNA fragments up to 1000 bp. Comparisons of electropherograms provided genomic fingerprint templates which could be further used for supplementary information. The ERIC-PCR method coupled to CE provides a rapid technique in differentiating bacterial spp., and may contribute relevant information in food-borne outbreak studies.

摘要

分子技术,即肠杆菌重复基因间共识序列(ERIC)-聚合酶链反应(PCR),可产生区分细菌种类和菌株的基因组DNA指纹图谱。该技术被应用于单核细胞增生李斯特菌的特征分析,单核细胞增生李斯特菌是一种重要的食源性病原体,与众多李斯特菌病病例有关。ERIC-PCR产生了所有单核细胞增生李斯特菌血清型和李斯特菌属的独特DNA指纹图谱模式。基因组DNA指纹图谱的分析是使用毛细管电泳(CE)完成的,这是一种替代传统琼脂糖凝胶法的技术。CE条件(电动进样、施加电压)的优化使得扩增的DNA片段分辨率高达1000 bp。电泳图的比较提供了基因组指纹模板,可进一步用于补充信息。ERIC-PCR方法与CE相结合,提供了一种快速区分细菌种类的技术,并可能为食源性疾病暴发研究提供相关信息。

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