Development and evaluation of an immunomagnetic separation-ELISA for the detection of Staphylococcus aureus thermostable nuclease in composite milk

Yazdankhah, Siamak Pour; Sølverød, Liv; Simonsen, Sigrid; Olsen, Egil

doi:10.1016/s0378-1135(99)00035-8

Cited by 28 publications

(16 citation statements)

References 28 publications

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“…Combined with IMS, many detection methods can be used as a second step, to quantify and/or identify the captured bacteria. Most of the time, this approach has been directly applied in combination with selective culture-based methods [5], or the beads have been used as solid support for enzyme-linked immunosorbent assay (ELISA) [6, 7]. In addition, identification of captured bacteria can be realized by PCR after an elution step [8–13].…”

Section: Introductionmentioning

confidence: 99%

Rapid and Specific Enrichment of Culturable Gram Negative Bacteria Using Non-Lethal Copper-Free Click Chemistry Coupled with Magnetic Beads Separation

et al. 2015

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Currently, identification of pathogenic bacteria present at very low concentration requires a preliminary culture-based enrichment step. Many research efforts focus on the possibility to shorten this pre-enrichment step which is needed to reach the minimal number of cells that allows efficient identification. Rapid microbiological controls are a real public health issue and are required in food processing, water quality assessment or clinical pathology. Thus, the development of new methods for faster detection and isolation of pathogenic culturable bacteria is necessary. Here we describe a specific enrichment technique for culturable Gram negative bacteria, based on non-lethal click chemistry and the use of magnetic beads that allows fast detection and isolation. The assimilation and incorporation of an analog of Kdo, an essential component of lipopolysaccharides, possessing a bio-orthogonal azido function (Kdo-N3), allow functionalization of almost all Gram negative bacteria at the membrane level. Detection can be realized through strain-promoted azide-cyclooctyne cycloaddition, an example of click chemistry, which interestingly does not affect bacterial growth. Using E. coli as an example of Gram negative bacterium, we demonstrate the excellent specificity of the technique to detect culturable E. coli among bacterial mixtures also containing either dead E. coli, or live B. subtilis (as a model of microorganism not containing Kdo). Finally, in order to specifically isolate and concentrate culturable E. coli cells, we performed separation using magnetic beads in combination with click chemistry. This work highlights the efficiency of our technique to rapidly enrich and concentrate culturable Gram negative bacteria among other microorganisms that do not possess Kdo within their cell envelope.

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Section: Introductionmentioning

confidence: 99%

Rapid and Specific Enrichment of Culturable Gram Negative Bacteria Using Non-Lethal Copper-Free Click Chemistry Coupled with Magnetic Beads Separation

et al. 2015

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“…Although 11 capsular polysaccharide types are described, only two types, types 5 and 8, are clinically relevant in that they are predominant among clinical infection isolates of varied geographic origin (Roghmann et al 2005;Verdier et al 2007). S. aureus produces an extracellular nuclease protein which is heat stable with activity retained after incubation at 97°C for 60 min (Yazdankhah et al 1999). The activity of the enzyme has been shown to be preserved for at least 10 months in various foods at temperatures of 5±37°C (Lachica et al 1972).…”

Section: Introductionmentioning

confidence: 99%

“…The activity of the enzyme has been shown to be preserved for at least 10 months in various foods at temperatures of 5±37°C (Lachica et al 1972). Thermonuclease activity has been used for the rapid and direct detection of S. aureus in blood cultures (Madison and Baselski 1983) and foods (Yazdankhah et al 1999). Staphylococcal hemolysins are identified as important virulence factors that contribute to bacterial invasion and escape from the host immune response (Da Silva et al 2005).…”

Section: Introductionmentioning

confidence: 99%

Detection of virulence-associated genes in Staphylococcus aureus isolated from bovine clinical mastitis milk samples in Guangxi

Yang

Liang

et al. 2012

Trop Anim Health Prod

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“…The separation of nucleic acids from their native complex mixtures is required to enable processes such as sequencing, amplification, hybridization and detection. Magnetic particles have been used for various immuoassays including fluorescence or enzyme immunoassays [43]. …”

Section: Bioapplicationsmentioning

confidence: 99%

Magnetic Biotransport: Analysis and Applications

Furlani

2010

Materials

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Magnetic particles are finding increasing use in bioapplications, especially as carrier particles to transport biomaterials such as proteins, enzymes, nucleic acids and whole cells etc. Magnetic particles can be prepared with biofunctional coatings to target and label a specific biomaterial, and they enable controlled manipulation of a labeled biomaterial using an external magnetic field. In this review, we discuss the use of magnetic nanoparticles as transport agents in various bioapplications. We provide an overview of the properties of magnetic nanoparticles and their functionalization for bioapplications. We discuss the basic physics and equations governing the transport of magnetic particles at the micro- and nanoscale. We present two different transport models: a classical Newtonian model for predicting the motion of individual particles, and a drift-diffusion model for predicting the behavior of a concentration of nanoparticles that takes into account Brownian motion. We review specific magnetic biotransport applications including bioseparation, drug delivery and magnetofection. We demonstrate the transport models via application to these processes.

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Development and evaluation of an immunomagnetic separation-ELISA for the detection of Staphylococcus aureus thermostable nuclease in composite milk

Cited by 28 publications

References 28 publications

Rapid and Specific Enrichment of Culturable Gram Negative Bacteria Using Non-Lethal Copper-Free Click Chemistry Coupled with Magnetic Beads Separation

Rapid and Specific Enrichment of Culturable Gram Negative Bacteria Using Non-Lethal Copper-Free Click Chemistry Coupled with Magnetic Beads Separation

Detection of virulence-associated genes in Staphylococcus aureus isolated from bovine clinical mastitis milk samples in Guangxi

Magnetic Biotransport: Analysis and Applications

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