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

Parsonnet, Jeffrey; Mills, Joan T.; Gillis, Z A; Pier, Gerald B.

doi:10.1128/jcm.22.1.26-31.1985

Cited by 34 publications

(12 citation statements)

References 25 publications

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“…The purified protein yields a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (10% polyacrylamide), as demonstrated by a silver stain described by Gorg et al (9), and is serologically identical to TSST-1 preparations from other laboratories. TSST-1 was assayed with a competitive, enzyme-linked immunosorbent assay (19). Pu- rified TSST-1 contained <0.0003 ,ug of lipopolysaccharide per ,ug of TSST-1, as determined by a standard Limulus amebocyte lysate assay (Pyrogent; M. A. Bioproducts, Walkersville, Md.).…”

mentioning

confidence: 99%

A rabbit model of toxic shock syndrome that uses a constant, subcutaneous infusion of toxic shock syndrome toxin 1

Parsonnet¹,

Gillis²,

Richter³

et al. 1987

Infect Immun

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122

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We have developed a rabbit model of toxic shock syndrome that uses a subcutaneous infusion pump to administer toxic shock syndrome toxin 1 (TSST-1). A dose of 150 ,ug, infused at a constant rate over a period of 7 days, resulted in a characteristic illness highlighted by fever, conjunctival hyperemia, cachexia, and lethargy. The illness was uniformly fatal, with a mean interval until death of 3.2 ± 0.4 days. Serial determinations of serum chemistries confirmed the multisystem nature of this illness. Rabbits developed profound hypocalcemia, with levels falling from 15.5 ± 0.2 to 7.6 ± 0.4 mg/dl under the influence of TSST-1. Blood urea nitrogen and creatinine rose dramatically, in the setting of oliguria or anuria. Serum glutamicpyruvic transaminase was the most reliable indicator of hepatic dysfunction, with the mean rising from 48 U/liter before administration of TSST-1 to 546 U/liter among rabbits surviving 2 days of the infusion. Creatine phosphokinase also rose dramatically in 10 of 16 rabbits. Rabbits demonstrated relative neutrophilia and lymphopenia as well as an increase in the partial thromboplastin time. Histopathologic examination demonstrated disease of multiple organs, particularly the liver, spleen, and lymph nodes, all of which demonstrated inflammation, thrombosis, hemorrhage, and erythrophagocytosis. The concurrent administration of prednisolone with TSST-1 prevented death in four of four rabbits and greatly lessened the morbidity. Rabbits were not protected from morbidity or mortality by the concurrent administration of polymyxin B. We believe that a constant, subcutaneous infusion of TSST-1 in rabbits provides a reproducible model for studying the pathogenesis of TSS.

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mentioning

confidence: 99%

A rabbit model of toxic shock syndrome that uses a constant, subcutaneous infusion of toxic shock syndrome toxin 1

Parsonnet¹,

Gillis²,

Richter³

et al. 1987

Infect Immun

Self Cite

122

View full text Add to dashboard Cite

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“…b Determined by a competitive ELISA (19). % Reduction = [1 -(observed serologic activity/serologic activity relative to that of whole toxin)] x 100.…”

Section: Resultsmentioning

confidence: 99%

“…1% bovine serum albumin-0.2% sodium azide-0.05% Tween 20 (pH 7.4). The samples were analyzed by a competitive enzyme-linked immunosorbent assay (ELISA) (19). The quantities of active material were computed (in nanograms per milliliter) from calibration curves generated with purified TSST-1 standards in the range of 1 to 60 ng/ml.…”

Section: Methodsmentioning

confidence: 99%

Identification of functional antigenic segments of toxic shock syndrome toxin 1 by differential immunoreactivity and by differential mitogenic responses of human peripheral blood mononuclear cells, using active toxin fragments

Edwin

Kass

1989

Infect Immun

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When toxic shock syndrome toxin 1 was subjected to papain hydrolysis, two serologically active fragments of 16.3 kilodaltons (16K fragment) and 12.4 kilodaltons (12K fragment) were generated, whereas a third fragment of 9.7 kilodaltons (1OK fragment) was inactive. The biologic activities of the fragments were evaluated in vitro by determining their ability to promote nonspecific proliferation of human peripheral blood mononuclear cells. The 12K fragment was significantly (P c 0.013) more stimulatory than the 16K fragment. When human peripheral blood mononuclear cells were preincubated for a period of 24 h with various concentrations of the 16K fragment, followed by incubation with a constant amount (2 x 10-2 ng/ml) of whole toxin, the level of DNA synthesis induced by the holotoxin was reduced by approximately 60% when compared with that of controls exposed to whole toxin alone. The 12K fragment did not demonstrate a similar blocking effect. Immunoblots of the toxic shock syndrome toxin 1 digest, which were exposed to monoclonal antibodies (MAbs) developed against native toxin, depicted the presence of two different antigenic regions (epitopes). One MAb, 8-5-7, which has been shown previously to inhibit the biologic activity of the holotoxin in vitro and in vivo, reacted primarily with the 12K fragment. A second MAb, 10-6-1, that did not neutralize interleukin-1 production reacted primarily with the 16K fragment. On the basis of the differential mitogenic responses and the identification of heterologous epitopes, it was concluded that the functional region of the holotoxin can be partitioned into at least two functional segments encompassed between amino acid residues 53 and 87 and between amino acid residues 88 and 194 on the polypeptide chain.

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“…Methods, such as radioimmunoassay (5) and enzymelinked immunosorbent assay (9), that are equally as sensitive as the RPLA method, have been developed for the assay of TSST-1. However, the disadvantages of the radioimmunoassay are the need for handling radioactive materials and the requirement for sophisticated equipment such as a scintillation counter.…”

Section: Discussionmentioning

confidence: 99%

Latex agglutination test for staphylococcal toxic shock syndrome toxin 1

Igarashi¹,

Fujikawa²,

Shingaki³

et al. 1986

J Clin Microbiol

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A reversed passive latex agglutination method, in which latex particles were sensitized with specific anti-toxic shock syndrome toxin-i (TSST-1) immunoglobulin, was found to be a simple and sensitive method for the detection of TSST-1 production by Staphylococcus aureus strains. The minimum amount of TSST-1 detectable was approximately 1.0 nglml. Of 41 S. aureus isolates from toxic shock syndrome patients and controls, 23 were positive for TSST-1 production, whereas only 20 strains were positive for TSST-1 production by an Ouchterlony immunodiffusion method. The reversed passive latex agglutination method was used to examine S. aureus strains isolated in Japan from staphylococcal infections, feces from healthy individuals, food from poisoning outbreaks, and market food.

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

Cited by 34 publications

References 25 publications

A rabbit model of toxic shock syndrome that uses a constant, subcutaneous infusion of toxic shock syndrome toxin 1

A rabbit model of toxic shock syndrome that uses a constant, subcutaneous infusion of toxic shock syndrome toxin 1

Identification of functional antigenic segments of toxic shock syndrome toxin 1 by differential immunoreactivity and by differential mitogenic responses of human peripheral blood mononuclear cells, using active toxin fragments

Latex agglutination test for staphylococcal toxic shock syndrome toxin 1

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