Subtyping of Listeria monocytogenes Based on Nucleotide Polymorphism in the clpC, inlA, hlyA, and plcA Genes and Rapid Identification of L. monocytogenes Genetically Similar to Clinical Isolates

To develop a new method for identification of Listeria monocytogenes genetically similar to clinical isolates, single-nucleotide polymorphism (SNP) typing and multi-locus sequence typing (MLST) of 126 isolates of L. monocytogenes from clinical and environmental samples were performed based on sequen...

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Published in:Food Science and Technology Research 2008, Vol.14(6), pp.557-557
Main Authors: HONJOH, Ken-ichi, FUJIHARA, Kumiko, HARAGUCHI, Takahiro, ONO, Yukari, KOBAYASHI, Hiroshi, HIWAKI, Hiroshi, KAMIKADO, Hideaki, JANG, Sung Sik, RYU, Sangryeol, MIYAMOTO, Takahisa
Format: Article
Language:English
Subjects:
Clinical isolates
cycling probe technology
Food
Gene polymorphism
Listeria monocytogenes
Phospholipase C
Polymerase chain reaction
Primers
Probes
real-time PCR
Serotypes
Single-nucleotide polymorphism
SNP typing
Typing
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Summary:To develop a new method for identification of Listeria monocytogenes genetically similar to clinical isolates, single-nucleotide polymorphism (SNP) typing and multi-locus sequence typing (MLST) of 126 isolates of L. monocytogenes from clinical and environmental samples were performed based on sequence analysis of parts of four genes (hlyA, clpC, inlA, and plcA). Based on the sequences of the isolates in this study, SNP typing showed that hlyA, clpC, inlA, and plcA genes were categorized into 9, 14, 17, and 21 types, respectively. MLST showed that the isolates were grouped into 35 types including 12 types of clinical isolates. Out of those, four MLST types were found in food or environmental and clinical isolates. In particular, all clinical isolates with serotype 1/2a were grouped into the same hlyA SNP A5 type. A method using real-time PCR combined with Cycling Probe Technology was developed for rapid identification of SNP type of L. monocytogenes genetically similar to the clinical isolates. By using this method, the 1/2a clinical isolates showing MLST-2 were successfully identified with a specific primer set and a cycling probe designed on the basis of sequence of hlyA. Furthermore, clinical isolates of serotype 4b showing MLST-4 or -35 were successfully identified by a method using cycling probes based on sequences of clpC and inlA.
ISSN:1344-6606
1881-3984
1881-3984
DOI:10.3136/fstr.14.557