Simultaneous quantitative detection of Listeria spp. and Listeria monocytogenes using a duplex real-time PCR-based assay

被引:4
|
作者
Rodríguez-Lázaro, D
Hernández, M
Pla, M [1 ]
机构
[1] Univ Girona, Inst Food & Agr Technol, INTEA, E-17071 Girona, Spain
[2] CSIC, Inst Mol Biol, IBMB, E-08034 Barcelona, Spain
关键词
Listeria spp; Listeria monocytogenes; real-time PCR; multiplex; food-borne pathogen detection;
D O I
10.1016/j.femsle.2004.02.018
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
We report a duplex real-time PCR-based assay for the simultaneous quantitative detection of Lisieria 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 Lisieria 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. (C) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码:257 / 267
页数:11
相关论文
共 50 条
  • [1] Molecular beacon based real-time PCR assay for simultaneous detection of Listeria monocytogenes and Salmonella spp. in dairy products
    Singh, Jitender
    Batish, Virender K.
    Grover, Sunita
    [J]. DAIRY SCIENCE & TECHNOLOGY, 2011, 91 (03) : 373 - 382
  • [2] Multiplex Real-time PCR for the Simultaneous Detection of Salmonella spp. and Listeria monocytogenes in Food Samples
    Olaya Ruiz-Rueda
    Marçal Soler
    Laia Calvó
    Jesús L. García-Gil
    [J]. Food Analytical Methods, 2011, 4 : 131 - 138
  • [3] Multiplex Real-time PCR for the Simultaneous Detection of Salmonella spp. and Listeria monocytogenes in Food Samples
    Ruiz-Rueda, Olaya
    Soler, Marcal
    Calvo, Laia
    Garcia-Gil, Jesus L.
    [J]. FOOD ANALYTICAL METHODS, 2011, 4 (02) : 131 - 138
  • [4] A novel real-time PCR-based method for the detection of Listeria monocytogenes in food
    Oravcova, K.
    Kuchta, T.
    Kaclikova, E.
    [J]. LETTERS IN APPLIED MICROBIOLOGY, 2007, 45 (05) : 568 - 573
  • [5] Development and validation of qualitative SYBR®Green Real-Time PCR for detection and discrimination of Listeria spp. and Listeria monocytogenes
    Barbau-Piednoir, Elodie
    Botteldoorn, Nadine
    Yde, Marc
    Mahillon, Jacques
    Roosens, Nancy H.
    [J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 2013, 97 (09) : 4021 - 4037
  • [6] Quantitative detection of Listeria monocytogenes in biofilms by real-time PCR
    Guilbaud, M
    de Coppet, P
    Bourion, F
    Rachman, C
    Prévost, H
    Dousset, X
    [J]. APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2005, 71 (04) : 2190 - 2194
  • [7] Development and validation of qualitative SYBR®Green Real-Time PCR for detection and discrimination of Listeria spp. and Listeria monocytogenes
    Elodie Barbau-Piednoir
    Nadine Botteldoorn
    Marc Yde
    Jacques Mahillon
    Nancy H. Roosens
    [J]. Applied Microbiology and Biotechnology, 2013, 97 : 4021 - 4037
  • [8] Detection and quantification of the iap gene of Listeria monocytogenes and Listeria innocua by a new real-time quantitative PCR assay
    Hein, I
    Klein, D
    Lehner, A
    Bubert, A
    Brandl, E
    Wagner, M
    [J]. RESEARCH IN MICROBIOLOGY, 2001, 152 (01) : 37 - 46
  • [9] Real-time PCRs assay for serogrouping Listeria monocytogenes and differentiation from other Listeria spp.
    Vitullo, Monia
    Grant, Kathie Ann
    Sammarco, Michela Lucia
    Tamburro, Manuela
    Ripabelli, Giancarlo
    Amar, Corinne Francoise Laurence
    [J]. MOLECULAR AND CELLULAR PROBES, 2013, 27 (01) : 68 - 70
  • [10] Comparison of three real-time PCR-based methods for the detection of Listeria monocytogenes in food
    Oravcova, Katarina
    Trncikova, Terezia
    Kaclikova, Eva
    [J]. JOURNAL OF FOOD AND NUTRITION RESEARCH, 2007, 46 (02): : 63 - 67