Detection and Differentiation of<i>Listeria</i>spp. by a Single Reaction Based on Multiplex PCR

Bubert, Andreas; Hein, Inge; Rauch, Marcus; Lehner, Angelika; Yoon, Byoung-Su; Goebel, Werner; Wagner, Martin

doi:10.1128/aem.65.10.4688-4692.1999

Cited by 190 publications

(99 citation statements)

References 20 publications

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“…Identification of bacteria in the mixed culture was performed using standard PCR using species-specific primers derived from the iap gene in Listeria (Bubert et al 1999): Lis1B (5¢-TTATACGCGACCGAAGCCAAC-3¢) and MonoA (5¢-CAAACTGCTAACACAGCTACT-3¢), Ino2 (5¢-ACTAGCACTCCAGTTGTTAAAC-3¢). For L. monocytogenes, a combination of primers Lis1B and MonoA (660 bp) was used, and Lis1B and Ino2 (870 bp) were used for L. innocua.…”

Section: Bacterial Identification By Pcrmentioning

confidence: 99%

Human heat-shock protein 60 receptor-coated paramagnetic beads show improved capture of Listeria monocytogenes in the presence of other Listeria in food

Koo

Aroonnual

Bhunia

2011

Journal of Applied Microbiology

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Aims: To investigate the suitability of human Hsp60, a receptor for Listeria adhesion protein (LAP), on paramagnetic beads (PMB) to capture Listeria monocytogenes from food in the presence of other Listeria to facilitate rapid and specific detection of this pathogen. Methods and Results: Commercially available streptavidin‐coated PMBs were linked with biotinylated Hsp60 (PMB‐Hsp60), and the bacterial capture efficiency from pure culture and meat samples was determined. Capture rate was also compared with the monoclonal antibody (MAb)‐C11E9‐coated beads (PMB‐C11E9) and the commercial Dynabeads anti‐Listeria. Captured cells were detected and quantified by plating on selective medium, quantitative real‐time PCR (qPCR) and a light‐scattering sensor. Overall, all ligand‐coated beads had similar capture efficiency (varied from 1·8 to 9·2%) for L. monocytogenes under the conditions employed, and the minimum cell number required to achieve such capture was 103 CFU ml−1. PMB‐Hsp60 had significantly greater capture efficiency for pathogenic Listeria (P < 0·0001) than the nonpathogenic Listeria. In contrast, PMB‐C11E9 and Dynabeads anti‐Listeria had similar capture efficiency for both. The efficacy of all PMBs to capture L. monocytogenes in the presence of Listeria innocua from food matrices was compared. Although Dynabeads anti‐Listeria had the overall best capture efficiency, PMB‐Hsp60 was able to selectively capture L. monocytogenes even in the presence of 10–100‐fold more L. innocua cells from enriched meat samples. Conclusions: Data show that the human cell receptor, Hsp60, is suitable for the capture of pathogenic Listeria on PMB in the presence of other Listeria in food. Significance and Impact of the Study: As pathogen interaction with host cells is highly specific, host cell receptors could be used as alternate capture molecules on PMB to aid in specific detection of pathogens.

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Section: Bacterial Identification By Pcrmentioning

confidence: 99%

Human heat-shock protein 60 receptor-coated paramagnetic beads show improved capture of Listeria monocytogenes in the presence of other Listeria in food

Koo

Aroonnual

Bhunia

2011

Journal of Applied Microbiology

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“…This infection is acquired via the consumption of contaminated, most often ready-to-eat, food [2][3][4]. L. monocytogenes usually co-exists with other species of this genus such as the frequent and generally non-pathogenic Listeria innocua, which can be used as an indicator of the possible presence of L. monocytogenes in food [5][6][7], or Listeria ivanovii and Listeria seeligeri, which occasionally cause human infections [8,9]. The food safety regulations of most countries tolerate no L. monocytogenes in ready-to-eat food [10], although the minimal infectious dose is generally thought to be higher than 100 viable cells [11].…”

Section: Introductionmentioning

confidence: 99%

“…Multiplex PCR allows the simultaneous amplification of more than one target sequence in a single reaction. This saves considerable time and effort, and decreases the number of reactions that need to be performed to detect food-borne pathogens in a food sample [5,[14][15][16]. Several conventional PCR assays targeting the Listeria genus and L. monocytogenes have been reported [5,[17][18][19][20].…”

Section: Introductionmentioning

confidence: 99%

“…This saves considerable time and effort, and decreases the number of reactions that need to be performed to detect food-borne pathogens in a food sample [5,[14][15][16]. Several conventional PCR assays targeting the Listeria genus and L. monocytogenes have been reported [5,[17][18][19][20]. Furthermore, single RTi-PCR assays for the detection of L. monocytogenes have been described [21][22][23][24][25].…”

Section: Introductionmentioning

confidence: 99%

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

Rodrı́guez-Lázaro

Hernández

Pla

2004

FEMS Microbiology Letters

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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 R2 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.

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“…It consisted of a mixture of stab cultures which had been inoculated during the 1980s and lyophils which had been prepared in the 1950s-1960s, These tubes had always been stored at ambient temperature until 2008, when they were transferred to University College Cork, where they have been stored at 10°C. A subset of the SLCC was deposited at the Institute Pasteur, Paris, and selected even smaller subsets have also been supplied to various laboratories (Loessner and Busse, 1990;Bubert et al, 1999;Friedly et al, 2008). However, most of the publicly available information regarding the SLCC only existed as handwritten or typed documents, largely in German.…”

Section: Introductionmentioning

confidence: 99%

Revival of Seeliger's historical ‘Special Listeria Culture Collection’

Haase

Murphy

Choudhury

et al. 2011

Environmental Microbiology

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The recent evolution of bacterial species can be elucidated with the aid of large historical strain collections. Unfortunately, information for some of these strain collections is not publicly available, or only in a format which is not readily digitized. The form of storage of traditional collection often requires considerable space and microbiological access to the individual strains can be time consuming. One such historical strain collection was assembled by Professor H.P. Seeliger, the so-called 'Special Listeria Culture Collection' (SLCC). The SLCC contains over 6000 Listeria strains which had been isolated between 1921 and 1987. The information on the properties of the strains was hand written or typed, primarily in German, and the stabs and lyophils used for storage were not ordered. Here we present a description of this strain collection after resuscitation and digitalization. Data were transcribed into a relational database and the revived bacterial strains were stored in a robotically friendly format, where the location of each tube is stored in a database. We resuscitated 4404 Listeria strains from the SLCC, and summarize their properties as well as making the detailed strain information publicly available. This digital information and the revival of the SLCC will facilitate historical analyses of the phylogeography of Listeria.

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Detection and Differentiation ofListeriaspp. by a Single Reaction Based on Multiplex PCR

Cited by 190 publications

References 20 publications

Human heat-shock protein 60 receptor-coated paramagnetic beads show improved capture of Listeria monocytogenes in the presence of other Listeria in food

Human heat-shock protein 60 receptor-coated paramagnetic beads show improved capture of Listeria monocytogenes in the presence of other Listeria in food

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

Revival of Seeliger's historical ‘Special Listeria Culture Collection’

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