PREVENTION OF CROSS‐REACTIONS IN THE ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) FOR THE DETECTION OF <i>STAPHYLOCOCCUS AUREUS</i> ENTEROTOXIN TYPE B IN CULTURE FILTRATES AND FOODS

Koper, Jan W.; Hagenaars, A. M.; Notermans, S.

doi:10.1111/j.1745-4565.1980.tb00389.x

Cited by 36 publications

(18 citation statements)

References 13 publications

(12 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This result was not in agreement with the findings of Hahn et al (1986) who found that the use of a biotin-rabbit IgG conjugate prevented interference due to protein A in an ELISA system for the detection of enterotoxins A and B. The binding to the Fc region of the IgGs of many species by protein A can lead to false positive results in i:mmunodetection (Koper et al 1980). That is the reason why some authors used sample treatments such as chromatography (Berdal et al 1981;Lapeyre et al 1987) or incubation with insoluble porcine IgG (Fey & Buckhard 1981).…”

Section: N D I R E C T S a N D W I C H E L I S Acontrasting

confidence: 72%

Protein A insensitive ELISA detection of staphylococcal enterotoxin B

Rogemond

Castro

Delaunay

et al. 1991

Lett Appl Microbiol

View full text Add to dashboard Cite

A sandwich enzyme-linked immunosorbent assay to detect staphylococcal enterotoxin B was developed using rat monoclonal antibodies as capture antibodies and as a biotinylated conjugate. This test was sensitive, less than 1 ng/ml of enterotoxin B was detected and interference by protein A was prevented by the use of rat monoclonal antibodies of the IgG2a isotype which were insensitive to protein A even at concentrations greater than 1000 ng/ml.

show abstract

Section: N D I R E C T S a N D W I C H E L I S Acontrasting

confidence: 72%

Protein A insensitive ELISA detection of staphylococcal enterotoxin B

Rogemond

Castro

Delaunay

et al. 1991

Lett Appl Microbiol

View full text Add to dashboard Cite

show abstract

“…Effect of detergents. The extraction and concentration of SE from the food sample is an essential and laborious step in any enzyme immunoassay (9,11,19,23,37). Since foods are complex systems where hydrophobic interactions are common (lipid-lipid, protein-protein, lipid-protein), detergents are likely to help minimize interference by these constituents reacting with the immunoreagents.…”

Section: Discussionmentioning

confidence: 99%

Rapid and sensitive sandwich enzyme-linked immunosorbent assay for detection of staphylococcal enterotoxin B in cheese

Morissette

Goulet

Lamoureux

1991

Appl Environ Microbiol

View full text Add to dashboard Cite

A rapid and sensitive screening sandwich enzyme-linked immunosorbent assay (ELISA) was developed for the detection of staphylococcal enterotoxin B (SEB) in cheese by using a highly avid anti-SEB antibody (Ab) as the capture Ab (CAb) and as the biotinylated Ab conjugate. The glutaraldehyde fixation method for the immobilization of CAb on polystyrene dipsticks was superior to the adsorption fixation and the adsorptionglutaraldehyde fixation methods. The glutaraldehyde fixation method resulted in a higher surface-saturating CAb concentration as evaluated by the peroxidase saturation technique and by the ability of the CAb-coated dipstick to discriminate between positive and negative controls (index of discrimination). Of nine blocking agents used alone or in pairs, lysine-human serum albumin, bovine serum albumin, human serum albumin, and gelatin effectively saturated available sites on the CAb-coated dipsticks without causing interference with the antigen-Ab reactions. The addition of 1% polyethylene glycol to the diluent of the biotinylated anti-SEB Ab conjugate improved the detection of SEB. A concentration of 4% polyethylene glycol allowed a 5-min reaction time for the streptavidin-biotin-horseradish peroxidase conjugate. Cheddar cheese homogenate reduced the sensitivity of the SEB assay; however, the sensitivity was restored when 1.6% (wt/vol) of either a nonionic detergent (Mega-9) or two zwitterionic detergents (Zwittergent 3-10 and 3-12 detergent) was added to the diluent. By using the rapid sandwich ELISA, a minimum of 0.5 to 1.0 ng of SEB per ml was detected within 45 min. The whole procedure for the analysis of the cheddar cheese samples was completed within 1 h. This sandwich ELISA could be a rapid and sensitive screening method for the detection of staphylococcal enterotoxins in food for the agri-food industries.

show abstract

“…ELISAs are being used with increasing frequency to measure a variety ofbacterial products. ELISAs are widely used for the quantitation of staphylococcal enterotoxins in culture fluids and food samples (1,7,8,12,13,19,22) as well as other staphylococcal products such as alpha-toxin (23) and protein A (6). The detection of bacterial antigens in staphylococcal culture supernatants by an ELISA, however, is complicated by thq presence of protein A.…”

Section: Discussionmentioning

confidence: 99%

Competitive, enzyme-linked immunosorbent assay for toxic shock syndrome toxin 1

Parsonnet¹,

Mills²,

Gillis³

et al. 1985

J Clin Microbiol

View full text Add to dashboard Cite

We developed a competitive, enzyme-linked immunosorbent assay for the quantitation of toxic shock syndrome toxin 1 (TSST-1). Polyvalent immunoglobulin G from immunized rabbits was used as the capture antibody, and alkaline phosphatase conjugated to purified toxin served as the indicator enzyme. A standard curve was-génerated with each experiment, from which the concentration of toxin in culture supernatahts was extrapolated. The assay was useful for determining toxin concentrations of 0.03 to 0.5 ,g/ml, which is a substantial, practical improvement over immunodiffusion methods.,Staphylococcal enterotoxins A through E were not significantly cross-reactive in the assay, and staphylococcal protein A did not interfere with quantitation of TSST-1. By testing a variety ot staphylococcal strains, we found 100% concordance between toxin determinations made with our assay and those made by the investigators from whom the strains were obtained. The competitive, enzyme-linked immunosorbent assay is a highly reproducible, inexpensive means of determining TSST-1 concentrations and may have broad applicability in the field of toxic shock research.

show abstract

PREVENTION OF CROSS‐REACTIONS IN THE ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) FOR THE DETECTION OF STAPHYLOCOCCUS AUREUS ENTEROTOXIN TYPE B IN CULTURE FILTRATES AND FOODS

Cited by 36 publications

References 13 publications

Protein A insensitive ELISA detection of staphylococcal enterotoxin B

Protein A insensitive ELISA detection of staphylococcal enterotoxin B

Rapid and sensitive sandwich enzyme-linked immunosorbent assay for detection of staphylococcal enterotoxin B in cheese

Competitive, enzyme-linked immunosorbent assay for toxic shock syndrome toxin 1

Contact Info

Product

Resources

About