一种快速鉴定单增李斯特菌的方法

A Rapid Method to Identify Listeria monocytogenes

通过单增李斯特菌的生化特性对李斯特菌进行鉴定,并根据单增李斯特菌的actA基因序列,以及其他李斯特菌的特异性靶序列设计引物,用PCR技术快速鉴定单增李斯特菌。结果表明,生化实验时间较长且误差较大,而通过PCR能成功扩增出827bp的特征性条带,同时用CTAB/NaCl法和热裂解法分别提取基因组后进行PCR和电泳,结果两种方法都可以很好的扩增出特征性条带,而热裂解法由于更简便快速,实验确定了热裂解法是一种快速有效的鉴定单增李斯特菌的方法。

Listeria spp. was identified acrroding to the biochemical characteristics ofListeria monocytogenes, and a polymerase chain reaction （PCR） assay based on the primes targeting the gene actA and other specific primes of Listeria spp. was developed for detecting Listeria monocytogenes. The results showed that biochemical assays are time-consuming and often make errors while 827 bp band was detected successfully by PCR amplification. The CTAB/NaCl and boiling method were respectively used to extract genome DNA, both them could amply the specific band after PCR and electrophoresis by detecting on agarose gel. The boiling method is a more simple and convenient one to identify Listeria monocytogenes by the comparison of CTAB/NaCl and it is effective to prepare the template DNA.