单核细胞增生李斯特菌的脉冲场凝胶电泳（PFGE）分析

Pulsed-field gel electrophoresis (PFGE) analysis of Listeria monocytogenes.

作者信息

Dalmasso Marion, Jordan Kieran

机构信息

Teagasc Food Research Centre, Moorepark, Fermoy, Co., Cork, Ireland.

出版信息

Methods Mol Biol. 2014;1157:63-72. doi: 10.1007/978-1-4939-0703-8_5.

DOI:10.1007/978-1-4939-0703-8_5

PMID:24792548

Abstract

PFGE is a valuable tool for assessing Listeria monocytogenes strain interrelatedness. It is based on the study of total bacterial DNA restriction patterns. Cells are embedded in agarose plugs before being lysed. The released DNA is then digested into large fragments by restriction enzymes. As DNA fragments are too large to be separated by traditional electrophoresis in an agarose gel, changes in the direction of the electrical current are periodically applied in order to allow the proper migration of large DNA fragments. Strains are characterized by the obtained DNA fragment patterns or pulsotypes which vary depending on the number and size of bands.

摘要

脉冲场凝胶电泳（PFGE）是评估单核细胞增生李斯特菌菌株相关性的一种重要工具。它基于对细菌总DNA限制性图谱的研究。细胞在裂解前被包埋在琼脂糖凝胶块中。然后，释放出的DNA被限制性内切酶消化成大片段。由于DNA片段太大，无法通过传统的琼脂糖凝胶电泳进行分离，因此需要周期性地改变电流方向，以使大片段DNA能够正确迁移。菌株通过所获得的DNA片段图谱或脉冲型进行特征分析，这些图谱或脉冲型会因条带的数量和大小而有所不同。