Development of a monoclonal antibody-based colony blot immunoassay for detection of thermotolerant Campylobacter species

Huang, Hongsheng; Phipps-Todd, Beverley; McMahon, Tanis; Elmgren, C.; Lutze-Wallace, Cyril; Todd, Zoe A.; García, Manuel

doi:10.1016/j.mimet.2016.08.015

Cited by 7 publications

(7 citation statements)

References 14 publications

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“…2B ), despite the introduction of numerous different molecular techniques for the detection of Campylobacter over the same time period 34 , 35 , 37 . Although C. hyointestinalis is considered to be a non-thermotolerant member of the Campylobacteraeceae family 33 , it has been isolated most commonly under selective conditions that exploit high temperatures (>41 °C) (Fig. 2C ), possibly because these are the prevailing conditions used for the identification of Campylobacter infections which are more commonly associated with the species C. jejuni and C. coli 38 .…”

Section: Resultsmentioning

confidence: 99%

“…Campylobacter hyointestinalis , originally isolated from diseased pigs 30 is a member of the “emerging Campylobacter spp.” group that can also cause disease in humans. Although C. hyointestinalis was originally described as being able to grow at 43 °C 31 , 32 it is frequently referred to as a non-thermotolerant species 33 . The species has two recognised subspecies; hyointestinalis and lawsonii .…”

Section: Introductionmentioning

confidence: 99%

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Updating the genomic taxonomy and epidemiology of Campylobacter hyointestinalis

Wilkinson

O’Donnell

Akhter

et al. 2018

Sci Rep

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Campylobacter hyointestinalis is a member of an emerging group of zoonotic Campylobacter spp. that are increasingly identified in both gastric and non-gastric disease in humans. Here, we discovered C. hyointestinalis in three separate classes of New Zealand ruminant livestock; cattle, sheep and deer. To investigate the relevance of these findings we performed a systematic literature review on global C. hyointestinalis epidemiology and used comparative genomics to better understand and classify members of the species. We found that C. hyointestinalis subspecies hyointestinalis has an open pangenome, with accessory gene contents involved in many essential processes such as metabolism, virulence and defence. We observed that horizontal gene transfer is likely to have played an overwhelming role in species diversification, favouring a public-goods-like mechanism of gene ‘acquisition and resampling’ over a tree-of-life-like vertical inheritance model of evolution. As a result, simplistic gene-based inferences of taxonomy by similarity are likely to be misleading. Such genomic plasticity will also mean that local evolutionary histories likely influence key species characteristics, such as host-association and virulence. This may help explain geographical differences in reported C. hyointestinalis epidemiology and limits what characteristics may be generalised, requiring further genomic studies of C. hyointestinalis in areas where it causes disease.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Updating the genomic taxonomy and epidemiology of Campylobacter hyointestinalis

Wilkinson

O’Donnell

Akhter

et al. 2018

Sci Rep

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“…E . faecalis colonies were analyzed for extracellular production of polyGlcNAc-polymers using a protocol previously described [97] with some modifications: Briefly, 0.45 μm nitrocellulose membranes (Bio-Rad) were placed on 1-day-old colonies grown on MOLP until they became completely wet. The plates/membranes were incubated at 37°C for 10 minutes prior to be carefully removed and transfer colony side up to a glass petri dish for air-drying (10 minutes at 37°C).…”

Section: Methodsmentioning

confidence: 99%

PolyGlcNAc-containing exopolymers enable surface penetration by non-motile Enterococcus faecalis

et al. 2019

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Bacterial pathogens have evolved strategies that enable them to invade tissues and spread within the host. Enterococcus faecalis is a leading cause of local and disseminated multidrug-resistant hospital infections, but the molecular mechanisms used by this non-motile bacterium to penetrate surfaces and translocate through tissues remain largely unexplored. Here we present experimental evidence indicating that E . faecalis generates exopolysaccharides containing β-1,6-linked poly- N -acetylglucosamine (polyGlcNAc) as a mechanism to successfully penetrate semisolid surfaces and translocate through human epithelial cell monolayers. Genetic screening and molecular analyses of mutant strains identified glnA , rpiA and epaX as genes critically required for optimal E . faecalis penetration and translocation. Mechanistically, GlnA and RpiA cooperated to generate uridine diphosphate N -acetylglucosamine (UDP-GlcNAc) that was utilized by EpaX to synthesize polyGlcNAc-containing polymers. Notably, exogenous supplementation with polymeric N -acetylglucosamine (PNAG) restored surface penetration by E . faecalis mutants devoid of EpaX. Our study uncovers an unexpected mechanism whereby the RpiA-GlnA-EpaX metabolic axis enables production of polyGlcNAc-containing polysaccharides that endow E . faecalis with the ability to penetrate surfaces. Hence, targeting carbohydrate metabolism or inhibiting biosynthesis of polyGlcNAc-containing exopolymers may represent a new strategy to more effectively confront enterococcal infections in the clinic.

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“…The protein based methods work either by immunological assays or gel electrophoresis. The immunological based techniques, such as ELISA (Enzyme Linked Immuno-Sorbant Assay), antigenic participation and agglutination assays and western blotting, which commonly are employed to identify microorganisms based on their surface epitopes, are very specific and all are limited to the presence of a specific antibody for the given protein as well as the sensitivity and specificity of the antigen-antibody reactions [ 160 , 161 ].…”

Section: Other Methodsmentioning

confidence: 99%

Microbiological Sensing Technologies: A Review

2018

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Microorganisms have a significant influence on human activities and health, and consequently, there is high demand to develop automated, sensitive, and rapid methods for their detection. These methods might be applicable for clinical, industrial, and environmental applications. Although different techniques have been suggested and employed for the detection of microorganisms, and the majority of these methods are not cost effective and suffer from low sensitivity and low specificity, especially in mixed samples. This paper presents a comprehensive review of microbiological techniques and associated challenges for bioengineering researchers with an engineering background. Also, this paper reports on recent technological advances and their future prospects for a variety of microbiological applications.

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Development of a monoclonal antibody-based colony blot immunoassay for detection of thermotolerant Campylobacter species

Cited by 7 publications

References 14 publications

Updating the genomic taxonomy and epidemiology of Campylobacter hyointestinalis

Updating the genomic taxonomy and epidemiology of Campylobacter hyointestinalis

PolyGlcNAc-containing exopolymers enable surface penetration by non-motile Enterococcus faecalis

Microbiological Sensing Technologies: A Review

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