Open AccessSimultaneous quantitative detection of Listeria spp. and Listeria monocytogenes using a duplex real‐time PCR‐based assay
Author(s) -
RodríguezLázaro David,
Hernández Marta,
Pla Maria
Publication year - 2004
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2004.tb09490.x
Subject(s) - listeria monocytogenes , listeria , biology , microbiology and biotechnology , detection limit , 23s ribosomal rna , pathogen , real time polymerase chain reaction , ribosomal rna , bacteria , chromatography , chemistry , gene , genetics , rna , ribosome
We report a duplex real‐time PCR‐based assay for the simultaneous quantitative detection of Listeria spp. and the food‐borne pathogen Listeria monocytogenes . The targets of this single tube reaction were the 23S rDNA and hly genes of Listeria spp. and L. monocytogenes , respectively. Our assay was efficient, 100% selective (i.e., it allowed accurate simultaneous identification of 52 L. monocytogenes and 120 Listeria spp. strains through the FAM‐labelled hly and the VIC‐labelled 23S rDNA probes, respectively); and had a detection limit of one target molecule in 100% (23S rDNA) and 56% ( hly ) of the reactions. Simultaneous quantification was possible along a 5‐log dynamic range, with an upper limit of 30 target molecules and R 2 values >0.995 in both cases. Our results indicate that this assay based on the amplification of the 23S rDNA gene can accurately quantify any mixture of Listeria species and simultaneously unambiguously quantify L. monocytogenes .