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Identification of high‐risk Listeria monocytogenes serotypes in lineage I (serotype 1/2a, 1/2c, 3a and 3c) using multiplex PCR
Author(s) -
Nho S.W.,
Abdelhamed H.,
Reddy S.,
Karsi A.,
Lawrence M.L.
Publication year - 2015
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12876
Subject(s) - serotype , listeria monocytogenes , multiplex polymerase chain reaction , subtyping , biology , microbiology and biotechnology , multiplex , lineage (genetic) , virology , polymerase chain reaction , primer (cosmetics) , gene , genetics , bacteria , chemistry , organic chemistry , computer science , programming language
Aims Using molecular subtyping techniques, Listeria monocytogenes is divided into three major phylogenetic lineages, and a multiplex PCR method can differentiate five L. monocytogenes subgroups: 1/2a‐3a, 1/2c‐3c, 1/2b‐3b‐7, 4b‐4d‐4e and 4a‐4c. In this study, we conducted genome comparisons and evaluated serotype‐associated genes for their utility as a multiplex PCR‐based method for distinguishing high‐risk serotypes 1/2a and 1/2c in lineage I from low‐risk serotypes 3a and 3c. Methods and Results Primer sets were developed that are specific for serotype 1/2c ( LMOSLCC2372_0308 ) and serotype 3a ( LMLG_0742 ). These primers were then tested in a multiplex format with primers specific for serotype 1/2a ( flaA ) to separate serotypes 1/2a, 1/2c, 3a and 3c using 25 strains of lineage I L. monocytogenes . Conclusions Here, for the first time, we report primers specific for L. monocytogenes serotype 1/2c and serotype 3a, and we demonstrate a multiplex PCR method for separating the four serotypes of lineage I L. monocytogenes . Significance and Impact of the Study The described multiplex PCR assay consistently showed successful separation of 1/2a and 1/2c strains from 3a and 3c strains. PCR is routinely performed in many diagnostic and epidemiologic investigations for L. monocytogenes , and these primers should increase the feasibility and accessibility of L. monocytogenes serotyping.