The development of an efficient and rapid enzyme linked fluorescent assay method for the detection of Listeria spp. from foods

Sewell, A. M.; Warburton, Donald W.; Boville, A.; Daley, E.; Mullen, K.A.E.

doi:10.1016/s0168-1605(02)00221-0

Cited by 33 publications

(17 citation statements)

References 16 publications

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“…In this respect, the use of diagnostic PCR as a confirmatory test for presumptive colonies or even as a confirmatory test after the immunoenzymatic assay would improve accuracy of positive results obtained by both culture and mini-VIDAS. Irrespective of the method applied, we found that meat products, mainly refrigerated raw meat products, showed the highest incidence of L. monocytogenes as previously reported (Karpísková et al, 2000;Sewell et al, 2003;Mena et al, 2004). Compared to other surveys on L. monocytogenes in food-stuffs, an occurrence of 27 % is not alarming since most of the food products that tested positive are presumably going to be cooked before consumption.…”

Section: Discussionsupporting

confidence: 69%

“…Detection of L. monocytogenes: PCR versus mini-VIDAS 119 (Vaz-Velho et al, 2000;Sewell et al, 2003). However, when analysing the samples case-by-case, we found 10 % of discordant results.…”

Section: Discussionmentioning

confidence: 70%

“…Among them, mini-VIDAS L. monocytogenes (LMO) (BioMérieux Vitek, Inc., Missouri, USA) is a fully automated instrumental system that uses fluorescent ELFA (Enzyme Linked Fluorescent Assay) technology for the detection of listerial antigens in food. It has been applied in different studies on the incidence of L. monocytogenes (Vaz-Velho et al, 2000;Sewell et al, 2003;Mena et al, 2004;Silbernagel et al, 2004;Handa et al, 2005). PCR detection methods are rapid, sensitive, specific as well as suitable for automation and have been proposed to replace the timeconsuming culture-based traditional techniques (Hill 1996;Malorny et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

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PCR Detection of Listeria monocytogenes in different Food Products compared with the mini-VIDAS LMO System and the Standard Procedure ISO 11290-1

Aznar

Solís²

2006

J. Verbr. Lebensm.

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The presence of Listeria monocytogenes in 225 natural samples, including different food types, was investigated by three methods: (i) culture-based standard procedure ISO 11290-1, (ii) mini-VIDAS (Vitek Immuno Diagnostic Assay System) LMO, an enzyme linked fluorescent assay (ELFA) commercially available, and (iii) PCR using a previously established procedure. Identification of isolates recovered from the standard method and mini-VIDAS, on Oxford and PALCAM selective plates, was carried out using the API-Lis system and also by PCR, using L. monocytogenes specific primers. In all, 65 samples (64 meat products and one smoked salmon) were positive with any of the three procedures assayed. Taking into account the results based on PCR identification, the standard culture-based method found 22 and the mini-VIDAS 23, while PCR detected 60 positive samples in a total of 225 food samples. For comparative purposes, a set of "known positive samples" was established as reference that included 33 natural samples from which L. monocytogenes isolates (identified by specific PCR) had been recovered. The mini-VIDAS and ISO 11290-1 methods were equally sensitive. Compared to them PCR was clearly the more accurate and efficient procedure for detection of L. monocytogenes in food. Moreover, it showed no false negative results. The PCR approach can be completed in 48 working hours, and because of its specificity might eventually be cheaper than the other two procedures. Consequently we recommend PCR for routine detection of L. monocytogenes in food. Zusammenfassung(Redaktion): 225 Lebensmittelproben wurden auf das Vorhandensein von Listeria monocytogenes mit Hilfe von drei verschiedenen Methoden untersucht: (i) durch die auf Kultivierung basierende Standard-Methode nach ISO 11290-1, (ii) durch mini-VIDAS (Vitek Immuno Diagnostic Assay System) LMO, einem Enzym-basierten Verfahren mittels Fluoreszenz-Nachweis (ELFA; im Handel erhältlich) und (iii) durch ein PCR-Verfahren, das jüngst von den Autoren etabliert worden ist. Die Identifikation von Isolaten durch Einsatz der Standard-oder der mini-VI-DAS-Methode wurde nachvollzogen durch Verwendung des API-Lis-Systems und auch durch die PCR unter Verwendung von L. monocytogenes-spezifischen Primern. Insgesamt wurden 65 Lebensmittelproben durch jede der drei Methoden als positiv identifiziert. Durch die Standard-Methode wurden 22, durch die mini-VIDAS 23 und durch die PCR 60 positive Proben unter den 225 Lebensmittelproben identifiziert. Zu Vergleichszwecken wurden Referenzproben (belastet mit L. monocytogenes) durch PCR positiv identifiziert.Das mini-VIDAS-und das Verfahren nach ISO 11290-1 waren von gleicher Sensitivität; aber im Vergleich zu ihnen war die PCR deutlich exakter und effizienter, um L. monocytogenes in Lebensmitteln nachzuweisen. Das Ergebnis des Nachweises mittels PCR kann nach 48 Arbeitsstunden vorliegen. Daher empfehlen die Autoren ihre PCR als Routine-Verfahren für den Nachweis von L. monocytogenes in Lebensmitteln.

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Section: Discussionsupporting

confidence: 69%

“…Detection of L. monocytogenes: PCR versus mini-VIDAS 119 (Vaz-Velho et al, 2000;Sewell et al, 2003). However, when analysing the samples case-by-case, we found 10 % of discordant results.…”

Section: Discussionmentioning

confidence: 70%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

PCR Detection of Listeria monocytogenes in different Food Products compared with the mini-VIDAS LMO System and the Standard Procedure ISO 11290-1

Aznar

Solís²

2006

J. Verbr. Lebensm.

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“…from infant formulas require enrichment culture (48 h) and subculture on selective media followed by phenotypic identification; the procedures take at least 5 days (FDA/CFSAN 2002). Although the PCR and ELISA methods offer efficient and quantitative approaches for the detection of the pathogenic microorganism, the requirement of equipments and timeconsuming makes them infeasible for on-site inspection and quarantine (Liu et al 2006;Bhagwat 2003;O'Connor 2003;Sewell et al 2003). A novel nucleic acid amplification method, namely, loop-mediated isothermal amplification (LAMP), is capable of amplifying DNA under isothermal conditions with high sensitivity and speed (Notomi et al 2000).…”

Section: Introductionmentioning

confidence: 99%

Development of a loop-mediated isothermal amplification assay for detection of Cronobacter spp. (Enterobacter sakazakii)

Liu

Fang

Zhang

et al. 2011

World J Microbiol Biotechnol

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Contamination of Cronobacter spp. (Enterobacter sakazakii) in infant formulas and other food products is a severe problem. Here a loop-mediated isothermal amplification (LAMP) assay was developed for rapidly detecting Cronobacter spp. in powdered infant formula. Sequences of 16S/23S rDNA internal intergenic spacer of Cronobacter spp. were used as the target template to design LAMP primers. The detection outcome can be evaluated by the white precipitate or the fluorescence intensity under ultraviolet irradiation, both visible to naked eyes. The sensitivity and specificity of the LAMP assay was further analyzed in comparison with that of regular PCR and real time quantitative PCR. The results showed that all of Cronobacter spp. strains display positive reaction to the detections while all of the non-Cronobacter spp. strains were negative, and that the LAMP assay exhibits a high sensitivity of 9.1 fg/μL (The sensitivity of regular PCR and real time quantitative PCR is 91 and 9.1 pg/μL, respectively.). The amplified reaction could be accomplished in about 1 h, with the results visible to naked eyes. Hence, the LAMP assay developed by this study can provide a rapid and simple approach for the detection of Cronobacter spp. in infant formula.

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“…Conventional culture methods for detection of pathogens are time-consuming to produce the results, thus increasing the risk of pathogen uptake or transmission. Over the past several years, a variety of rapid methods have been investigated for detecting pathogens, including typical or derived immunological assays (Sewell et al 2003), nucleic acid-based tests (Amagliani et al 2004), and physicochemical tests based on bacterial growth (Firsten-Eden and Shelef 2000). These rapid methods have made significant progress towards reducing the total assay time.…”

mentioning

confidence: 99%

Rapid detection of pathogens using antibody-coated microbeads with bioluminescence in microfluidic chips

Guan

Zhang

et al. 2010

Biomed Microdevices

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Detection of pathogens was demonstrated in a polydimethylsiloxane (PDMS)/glass microfluidic chip with which microbead-based immunoseparation platform and the bioluminescence technology were integrated. Escherichia coli (E. coli) O157:H7 was used as the model bacteria. The microchamber in microfluidic chip was filled with glass beads coated with antibodies which could capture specific organism, and the capture efficiency of the chip for the bacteria was about 91.75% approximately 95.62%. Then the concentration of bacteria was determined by detecting adenosine triphosphate (ATP) employing bioluminescence reaction of firefly luciferin-lucifera-ATP on chip. The method allowed reliable detection of E. coli O157:H7 concentrations from 3.2 x 10(1) cfu/microL to 3.2 x 10(5) cfu/microL within 20 min. This research demonstrated excellent reproducibility, stability, and specificity, and could accurately detect the pathogenic bacteria in food samples. The microfluidic chip and the equipments used in this method are easy to miniaturize, thus the method has great potential to be developed to a portable device for rapid detection of pathogens.

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The development of an efficient and rapid enzyme linked fluorescent assay method for the detection of Listeria spp. from foods

Cited by 33 publications

References 16 publications

PCR Detection of Listeria monocytogenes in different Food Products compared with the mini-VIDAS LMO System and the Standard Procedure ISO 11290-1

PCR Detection of Listeria monocytogenes in different Food Products compared with the mini-VIDAS LMO System and the Standard Procedure ISO 11290-1

Development of a loop-mediated isothermal amplification assay for detection of Cronobacter spp. (Enterobacter sakazakii)

Rapid detection of pathogens using antibody-coated microbeads with bioluminescence in microfluidic chips

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