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样品处理对用于空肠弯曲菌鉴定的聚合酶链反应的影响。

Sample processing effect on polymerase chain reaction used for identification of Campylobacter jejuni.

作者信息

Sabatková Z, Pazlarová J, Demnerová K

机构信息

Department of Biochemistry and Microbiology, Institute of Chemical Technology, 166 28 Prague 6, Czechia.

出版信息

Folia Microbiol (Praha). 2004;49(6):693-7. doi: 10.1007/BF02931551.

Abstract

Model samples of Campylobacter jejuni for polymerase chain reaction (PCR) were prepared by rapid and simple procedures consisting of centrifugation, proteinase K treatment, Chelex 100 treatment, and boiling lyses. A PCR based on specific amplification of the variable sequence of 16S rRNA gene was performed using Tth DNA polymerase and the PCR products were visualized by agarose gel electrophoresis. The assay allowed the detection of 10 CFU/mL C. jejuni in the physiological saline and 100 CFU/mL in the basic Park and Sanders broth.

摘要

空肠弯曲菌聚合酶链反应(PCR)的模型样本通过快速简单的程序制备,包括离心、蛋白酶K处理、Chelex 100处理和煮沸裂解。使用Tth DNA聚合酶进行基于16S rRNA基因可变序列特异性扩增的PCR,并通过琼脂糖凝胶电泳观察PCR产物。该检测方法能够检测出生理盐水中10 CFU/mL的空肠弯曲菌以及基础帕克和桑德斯肉汤中100 CFU/mL的空肠弯曲菌。

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