Detection of the Staphylococcal Toxins

Bergdoll, Merlin S.

doi:10.1007/978-1-4613-0361-9_38

Cited by 12 publications

(7 citation statements)

References 14 publications

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Section: Introductionmentioning

confidence: 99%

“…Immunochemical assays were developed and utilized in the last 20 years for the detection of SE. The principles and effectiveness of these methods are described by Bergdoll (1996), Park et al (1992Park et al ( , 1994 and Wieneke (1991). Su and Wong (1997) also reviewed the biological, immunological and polymerase chain reaction-(PCR)-based methods for the detection of SE.…”

Section: Introductionmentioning

confidence: 99%

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A BIOSENSOR METHOD FOR A COMPETITIVE IMMUNOASSAY DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN B (SEB) IN MILK²

Medina

2005

Rapid Methods Automation Mic

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A sensitive and more rapid biosensor method for the detection of staphylococcal enterotoxins (SE) is needed by the food industry. Staphylococcal enterotoxin B (SEB) is highly heat resistant and is a potential bioterrorism agent. Our research objective was to develop a competitive immunoassay using a surface plasmon resonance (SPR) biosensor for the detection of SEB below 1 ng/mL [part per billion (ppb)] in fresh fluid milk. The assay consisted of SEB immobilization on the sensor surface. An anti‐SEB was allowed to bind with the SEB in samples off line prior to the biosensor analysis. The excess and unbound anti‐SEB was then captured by SEB sensor. The assay conditions were optimized to detect SEB in HEPES buffer and in whole milk. An analysis of milk samples spiked with 0.312–50 ppb. SEB consisted of heating the samples at 95C followed by rapid cooling and centrifugation at 2961 × g to separate the skim fraction. Aliquots of the skim fraction containing SEB were allowed to bind with anti‐SEB for 30 or 60 min. The SEB and anti‐SEB complex were separated from the free anti‐SEB by centrifugation, and the supernatants were injected over the sensor. SEB was detectable in buffer at 0.78–50 ppb and in spiked whole and skim milk from 0.312–25 ppb. The biosensor analysis including the sensor regeneration was 15 min per sample in a fully automated system. The competitive assay format resulted in higher detection sensitivity and greater sample throughput than the SPR biosensor sandwich assay. The competitive assay will be utilized for the detection of SEB in various foods and will be optimized for the detection of other staphylococcal toxins in foods.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A BIOSENSOR METHOD FOR A COMPETITIVE IMMUNOASSAY DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN B (SEB) IN MILK²

Medina

2005

Rapid Methods Automation Mic

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“…Klasična mikrobiološka diagnostika temelji na tem, da po videzu ločujemo različne bakterijske kolonije in iz njih delamo čiste kulture. O uspešnosti izolacije stafilokokov je v literaturi malo podatkov (18). V tem izbruhu smo tudi iz izbruhanine obolelega otroka uspeli izolirati bakterije S. aureus, verjetno zato, ker je bilo v zaužiti hrani veliko bakterij oz.…”

Section: Razpravaunclassified

Managing of the outbreak of staphylococcal food poisoning in primary school

Košnik¹,

Lah²,

Dermota³

et al. 2014

Slovenian Journal of Public Health

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aureus iz živila in humanih vzorcev smo opravili tipizacijo izolatov s pulzno elektroforezo (PFGE). Rezultati: Obolelo je 73 oseb od 374 izpostavljenih. Stopnja obolevnosti je bila 19,5-odstotna. Najpogostejši bolezenski znaki so bili: bruhanje (87,7 %), bolečine v trebuhu (75,3 %), driska (64,2 %) in slabost (59,3 %). Največje relativno tveganje (RT) za zastrupitev je bilo povezano z uživanjem mesnega sira (RT= 24,2 (95 % CI 12,5; p<0,001) in krompirjeve solate (RT= 19,4 (95 % CI 10,2; p<0,001 (87,7 %), stomach ache (75,3 %), diarrhea (64,2 %) and nausea ( 59,3 %). The highest relative risk (RR) were found for the meatloaf (RR= 24,2 (95 % CI 12,5; p<0,001)) and potato salad (RR= 19,4 (95 % CI 10,2; p<0,001)). Enterotoxin A producing S. aureus was isolated from hand of a cook, from potato salad and meatlof, from vomit and stools of patients. These strains had the same antibiotic sensitivity and were genetically closely related (96,3 %).

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“…The principles and performance of these immunological methods were described by Park et al. (1994), Bergdoll (1996) and Su and Wong (1997). The biological and polymerase chain reaction (PCR)‐based methods were also reviewed by Su and Wong (1997).…”

Section: Introductionmentioning

confidence: 99%

DEVELOPING A FLUORESCENT LATEX MICROPARTICLE IMMUNOASSAY USING ALEXA FLUOR 568 FOR DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN A¹

Medina¹

2007

Rapid Methods Automation Mic

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Staphylococcus aureus produces heat-stable toxins that cause major foodborne gastroenteritis. Staphylococcal enterotoxin A (SEA) is the most recovered toxin in staphylococcal food poisoning outbreaks. A sensitive method for the detection of SEA is needed for food safety and food defense monitoring. The objectives of this study were to develop a fluorescent immunoassay for the detection of SEA below the toxic level of 1 ng/mL and to minimize sample preparation. An affinity-purified anti-SEA was covalently linked to polystyrene microparticles. The SEA toxin was labeled with Alexa Fluor 568 (A568). The concentrations of these reagents were optimized for the detection of SEA in buffer and spiked hotdogs. SEA in samples was captured by the anti-SEA linked to the latex, followed by the binding of the SEA-A568 tracer. The latex-anti-SEA complex, bound with SEA and SEA-A568, was separated by centrifugation, and the fluorescent density of the supernatant was measured. SEA was detected at 0.25-10 ng/mL in buffer and in spiked hotdogs (ng/g). This fluorescent latex particle immunoassay can be utilized for the detection of SEA in a multitoxin detection system. PRACTICAL APPLICATIONSRapid and inexpensive methods are needed to detect trace levels of enterotoxins produced by Staphylococcus aureus organisms. Methods with 1 Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. 2 Corresponding 33 detection at or below 1 ng/g or 1 ng/mL are desired in testing food systems for food safety and food defense monitoring. Fluorescent immunoassays have the potential of providing high throughput analysis and increased sensitivity compared with enzyme immunoassays. The fluorescent latex microparticle immunoassay (FLMIA) using Alexa Fluor 568 as SEA antigen label detected SEA at 0.25-10 ng/mL in buffer solutions and 0.5 ng/g in spiked hot dogs. The FLMIA can be used for trace analysis in foods. This research shows the use of a fluorescent-labeled antigen as opposed to the use of labeled antibody (IgG). This research has shown the feasibility of using a long-wavelength fluorescent dye (Alexa Fluor 568) in immunodiagnostic assays. The use of the red fluorescent label for SEA will allow a multitoxin assay with green fluorescentlabeled SEB (staphylococcal enterotoxin B) in a single sample. 34 M.B. MEDINA 36 M.B. MEDINA

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Detection of the Staphylococcal Toxins

Cited by 12 publications

References 14 publications

A BIOSENSOR METHOD FOR A COMPETITIVE IMMUNOASSAY DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN B (SEB) IN MILK²

A BIOSENSOR METHOD FOR A COMPETITIVE IMMUNOASSAY DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN B (SEB) IN MILK²

Managing of the outbreak of staphylococcal food poisoning in primary school

DEVELOPING A FLUORESCENT LATEX MICROPARTICLE IMMUNOASSAY USING ALEXA FLUOR 568 FOR DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN A¹

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Detection of the Staphylococcal Toxins

Cited by 12 publications

References 14 publications

A BIOSENSOR METHOD FOR A COMPETITIVE IMMUNOASSAY DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN B (SEB) IN MILK2

A BIOSENSOR METHOD FOR A COMPETITIVE IMMUNOASSAY DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN B (SEB) IN MILK2

Managing of the outbreak of staphylococcal food poisoning in primary school

DEVELOPING A FLUORESCENT LATEX MICROPARTICLE IMMUNOASSAY USING ALEXA FLUOR 568 FOR DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN A1

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A BIOSENSOR METHOD FOR A COMPETITIVE IMMUNOASSAY DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN B (SEB) IN MILK²

A BIOSENSOR METHOD FOR A COMPETITIVE IMMUNOASSAY DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN B (SEB) IN MILK²

DEVELOPING A FLUORESCENT LATEX MICROPARTICLE IMMUNOASSAY USING ALEXA FLUOR 568 FOR DETECTION OF STAPHYLOCOCCAL ENTEROTOXIN A¹