Identification of six<i>Listeria</i>species by real-time PCR assay

Hage, E; Mpamugo, O; Ohai, C; Swift, Craig; Wooldridge, David; Amar, Corinne

doi:10.1111/lam.12223

Cited by 19 publications

(13 citation statements)

References 37 publications

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“…For rapid detection of L. monocytogenes in food matrices, lateral flow enzyme immunochromatography together with an immunomagnetic separation has been developed recently (Cho & Irudayaraj 2013). Molecular tools such as PCR, multiplex PCR and realtime PCR employing virulence-associated genes such as the mpl gene, prfA gene (Rossmanith et al 2006) and ssrA gene (O'grady et al 2008) have been found rapid, specific, reproducible and reliable (Portnoy et al 2002;Gianfranceschi et al 2013;Hage et al 2014;Khan et al 2014). Several multiplex PCR assays have been developed for simultaneous detection of various food-borne pathogens like Salmonella, Escherichia coli, Staphylococcus and also Listeria (Park et al 2006;Kawasaki et al 2009;Lee et al 2014).…”

Section: Diagnosismentioning

confidence: 98%

Listeriosis in animals, its public health significance (food-borne zoonosis) and advances in diagnosis and control: a comprehensive review

Dhama

Karthik

Tiwari

et al. 2015

Veterinary Quarterly

130

108

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Listeriosis is an infectious and fatal disease of animals, birds, fish, crustaceans and humans. It is an important food-borne zoonosis caused by Listeria monocytogenes, an intracellular pathogen with unique potential to spread from cell to cell, thereby crossing blood-brain, intestinal and placental barriers. The organism possesses a pile of virulence factors that help to infect the host and evade from host immune machinery. Though disease occurrence is sporadic throughout the world, it can result in severe damage during an outbreak. Listeriosis is characterized by septicaemia, encephalitis, meningitis, meningoencephalitis, abortion, stillbirth, perinatal infections and gastroenteritis with the incubation period varying with the form of infection. L. monocytogenes has been isolated worldwide from humans, animals, poultry, environmental sources like soil, river, decaying plants, and food sources like milk, meat and their products, seafood and vegetables. Since appropriate vaccines are not available and infection is mainly transmitted through foods in humans and animals, hygienic practices can prevent its spread. The present review describes etiology, epidemiology, transmission, clinical signs, post-mortem lesions, pathogenesis, public health significance, and advances in diagnosis, vaccines and treatment of this disease. Special attention has been given to novel as well as prospective emerging therapies that include bacteriophage and cytokine therapy, avian egg yolk antibodies and herbal therapy. Various vaccines, including advances in recombinant and DNA vaccines and their modes of eliciting immune response, are also discussed. Due focus has also been given regarding appropriate prevention and control strategies to be adapted for better management of this zoonotic disease.

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Section: Diagnosismentioning

confidence: 98%

Listeriosis in animals, its public health significance (food-borne zoonosis) and advances in diagnosis and control: a comprehensive review

Dhama

Karthik

Tiwari

et al. 2015

Veterinary Quarterly

130

108

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“…Furthermore, a novel 5′ exonuclease multiplex qPCR assay for the identification of six Listeria sp. including L. monocytogenes , L. seeligeri , L. ivanovii , L. welshimeri , L. grayi , and L. innocua was developed by Hage et al (2014) . In this study, two sets of triplex PCR were designed with one set identifying L. seeligeri , L. welshimeri , and L. monocytogenes and another set identifying L. ivanovii , L. grayi , and L. innocua .…”

Section: Molecular Detection Of Listeria Monocytogenesmentioning

confidence: 99%

An insight into the isolation, enumeration, and molecular detection of Listeria monocytogenes in food

et al. 2015

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Listeria monocytogenes, a foodborne pathogen that can cause listeriosis through the consumption of food contaminated with this pathogen. The ability of L. monocytogenes to survive in extreme conditions and cause food contaminations have become a major concern. Hence, routine microbiological food testing is necessary to prevent food contamination and outbreaks of foodborne illness. This review provides insight into the methods for cultural detection, enumeration, and molecular identification of L. monocytogenes in various food samples. There are a number of enrichment and plating media that can be used for the isolation of L. monocytogenes from food samples. Enrichment media such as buffered Listeria enrichment broth, Fraser broth, and University of Vermont Medium (UVM) Listeria enrichment broth are recommended by regulatory agencies such as Food and Drug Administration-bacteriological and analytical method (FDA-BAM), US Department of Agriculture-Food and Safety (USDA-FSIS), and International Organization for Standardization (ISO). Many plating media are available for the isolation of L. monocytogenes, for instance, polymyxin acriflavin lithium-chloride ceftazidime aesculin mannitol, Oxford, and other chromogenic media. Besides, reference methods like FDA-BAM, ISO 11290 method, and USDA-FSIS method are usually applied for the cultural detection or enumeration of L. monocytogenes. most probable number technique is applied for the enumeration of L. monocytogenes in the case of low level contamination. Molecular methods including polymerase chain reaction, multiplex polymerase chain reaction, real-time/quantitative polymerase chain reaction, nucleic acid sequence-based amplification, loop-mediated isothermal amplification, DNA microarray, and next generation sequencing technology for the detection and identification of L. monocytogenes are discussed in this review. Overall, molecular methods are rapid, sensitive, specific, time- and labor-saving. In future, there are chances for the development of new techniques for the detection and identification of foodborne with improved features.

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“…Listeria spp. are small Gram+, non spore-forming, facultative anaerobic bacteria widely distributed in the environment [ 12 - 14 ]. The Listeria genus consists of 10 species: Listeria monocytogenes, Listeria ivanovii, Listeria fleischmannii, Listeria innocua, Listeria welshimeri, Listeria seeligeri , Listeria martini, Listeria rocourtiae, Listeria weihenstephanensis and Listeria grayi [ 15 - 20 ].…”

Section: Introductionmentioning

confidence: 99%

Defensins from the tick Ixodes scapularis are effective against phytopathogenic fungi and the human bacterial pathogen Listeria grayi

et al. 2014

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BackgroundIxodes scapularis is the most common tick species in North America and a vector of important pathogens that cause diseases in humans and animals including Lyme disease, anaplasmosis and babesiosis. Tick defensins have been identified as a new source of antimicrobial agents with putative medical applications due to their wide-ranging antimicrobial activities. Two multigene families of defensins were previously reported in I. scapularis. The objective of the present study was to characterise the potential antimicrobial activity of two defensins from I. scapularis with emphasis on human pathogenic bacterial strains and important phytopathogenic fungi.MethodsScapularisin-3 and Scapularisin-6 mature peptides were chemically synthesised. In vitro antimicrobial assays were performed to test the activity of these two defensins against species of different bacterial genera including Gram-positive bacteria Staphylococcus aureus, Staphylococcus epidermidis, and Listeria spp. as well as Gram-negative bacteria Escherichia coli, Pseudomonas aeruginosa along with two plant-pathogenic fungi from the genus Fusarium. In addition, the tissue-specific expression patterns of Scapularisin-3 and Scapularisin-6 in I. scapularis midgut, salivary glands and embryo-derived cell lines were determined using PCR. Finally, tertiary structures of the two defensins were predicted and structural analyses were conducted.ResultsScapularisin-6 efficiently killed L. grayi, and both Scapularisin-3 and Scapularisin-6 caused strong inhibition (IC50 value: ~1 μM) of the germination of plant-pathogenic fungi Fusarium culmorum and Fusarium graminearum. Scapularisin-6 gene expression was observed in I. scapularis salivary glands and midgut. However, Scapularisin-3 gene expression was only detected in the salivary glands. Transcripts from the two defensins were not found in the I. scapularis tick cell lines ISE6 and ISE18.ConclusionOur results have two main implications. Firstly, the anti-Listeria and antifungal activities of Scapularisin-3 and Scapularisin-6 suggest that these peptides may be useful for (i) treatment of antibiotic-resistant L. grayi in humans and (ii) plant protection. Secondly, the antimicrobial properties of the two defensins described in this study may pave the way for further studies regarding pathogen invasion and innate immunity in I. scapularis.

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Identification of sixListeriaspecies by real-time PCR assay

Cited by 19 publications

References 37 publications

Listeriosis in animals, its public health significance (food-borne zoonosis) and advances in diagnosis and control: a comprehensive review

Listeriosis in animals, its public health significance (food-borne zoonosis) and advances in diagnosis and control: a comprehensive review

An insight into the isolation, enumeration, and molecular detection of Listeria monocytogenes in food

Defensins from the tick Ixodes scapularis are effective against phytopathogenic fungi and the human bacterial pathogen Listeria grayi

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