Rapid Detection of Gram-Negative Bacteriuria by Use of the Limulus Endotoxin Assay

Jorgensen, James H.; Carvajal, Hugo F.; Chipps, Bradley E.; Smith, Rodney F.

doi:10.1128/aem.26.1.38-42.1973

Cited by 57 publications

(25 citation statements)

References 9 publications

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“…While limulus activity has been used extensively as a measure of pathogenic potential in \arious infectious diseases (Ross et al 1975, Jorgensen & Lee 1978, it has also been used to determine differences in plaque pathogenicity (Fine et al 1978). Of interest is the finding that limulus activity on a ng/m!…”

Section: Discussionmentioning

confidence: 99%

The effect of rinsing with Listerine antiseptic on the properties of developing dental plaque

Fine

Letizia

Mandel

1985

J Clinic Periodontology

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Plaque was eoliected from a group of volunteers who used either Listerine antiseptic (LA), its vehicle control (V), or a water control (C) twiee daily in addition to their normal toothbrushing in a double-blind controlled clinical experiment. Following the 9-month clinical study, plaque collected from the supragingival surfaces of 20 teeth from each of 78 subjects was weighed wet, freeze dried, reweighed, resuspended, sonicated and estimated for protein. In addition, endotoxin activity was evaluated by means of the limulus lysate assay. A 52.6% reduction m wet weight was found LA versus C (/i = 0.04); LA versus V showed a 55.1% reduction (p = 0.03). A 59.0% reduction in dry weight was found LA versus C (/i = O.OI); LA versus V showed a 59.6% reduction (/J = 0.01). A 59.7% reduction in plaque protein was seen LA versus C (/J = 0.01); LA versus V showed a 59.2% decrease (p -O.Ol). A 75.8% reduction in limulus lysate activity was found LA versus C (p = O.0I); LA versus V showed a decrease of 77.9%i {p = 0.0\). Our results demonstrate that LA has a dramatic effect on plaque toxic activity, as measured by a decrease in limulus lysate assay, as well as on its biomass.

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Section: Discussionmentioning

confidence: 99%

The effect of rinsing with Listerine antiseptic on the properties of developing dental plaque

Fine

Letizia

Mandel

1985

J Clinic Periodontology

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“…In previous investigations, limulus activity in cementum-derived material from periodontally diseased teeth has been attributed to endotoxin-like material, with little further definition of the limulus-activating agents (Aleo et al 1974, Jones & O'Leary 1978. Indeed, many reports in different areas of research continue to equate limulus activity solely with levels of endotoxin (Ross et al 1975, Jorgensen & Lee 1978.…”

Section: Discussionmentioning

confidence: 99%

Preliminary characterization of material eluted from the roots of periodontally diseased teeth

Fine

Morris

Tabak

et al. 1980

J of Periodontal Research

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Eighteen periodontally diseased teeth and eleven normal teeth requiring surgical removal were each treated with one of four different solutions to elute potentially toxic material from their root surfaces. The solutions used were pyrogen‐free water, phenol‐water, trichloroacetic acid and citric acid. To define the composition of the eluted material, samples were analyzed for calcium, for ketodeoxyoctonic acid (KDO), a carbohydrate unique to Gram‐negative bacteria, and for limulus lysate activity. Limulus‐positive material was then analyzed for protein and nucleic acids and subjected to heat treatment, to the enzymes RNAse A and T1, and to lysozyme. It was found that TCA and citric acid removed more calcium and more toxic material from the root surface and subsurface than did either water or phenol‐water. The material extracted from 13 of the 18 diseased roots was limulus‐positive. Four of the limulus‐positive samples were also KDO‐positive, strongly indicating the presence of endotoxin in these samples. Neither these nor any of the other limulus‐positive responses were reversed or significantly reduced by exposure of samples to heat, to RNAse A and T1, or to lysozyme. In addition, limulus‐positive responses were unrelated to the amounts of protein and nucleic acids in the extracted material. When considered together, these preliminary characterizations indicate the presence of endotoxin in the material eluted from periodontally diseased teeth.

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“…Subsequent studies by Levin and co-workers established that the LAL assay could detect as little as 5 ng of circulating endotoxin per ml in plasma, thereby suggesting the potential utility of this assay in the detection of endotoxemia (15). Although the LAL assay has been proven clinically accurate in the detection of gram-negative bacteria (8) and meningitis (9), the most widespread use of the test has been in the pharmaceutical industry for in-process and end product pyrogen testing. Numerous products have been successfully evaluated for pyrogenicity by the LAL assay, including medical devices (25, 40), large-volume parenterals (25), small-volume parenterals (16), plasma fractions (23), and radiopharmaceuticals (2).…”

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confidence: 99%

Influence of fine structure of lipid A on Limulus amebocyte lysate clotting and toxic activities

Takayama¹,

Qureshi²,

Raetz³

et al. 1984

Infect Immun

101

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We examined the relationship between the fine structure of lipid A and the toxicity of endotoxin or lipopolysaccharides as measured by the Limulus amebocyte lysate (LAL), rabbit pyrogenicity, chicken embryo lethal dose, and dermal Shwartzman reaction tests. Lipid A and lipid Alike compounds obtained from deeprough mutants of Salmonella spp. and Escherichia coli had a wide range of structural variations. These compounds included native lipopolysaccharides, diphosphoryl and monophosphoryl lipid A's, and lipid X (a monosaccharide). The LAL test was positive for all lipids tested with lysates from Travenol Laboratories and from Associates of Cape Cod (2.9 x 103 to 2.6 x 107 endotoxin units per mg), except for O-deacylated and dephosphorylated lipid X, which were negative. The Mallinckrodt lysate gave negative tests for lipid X. In the rabbit pyrogenicity and chicken embryo lethal dose tests, only native lipopolysaccharide and diphosphoryl lipid A's were judged toxic. The Shwartzman reaction was positive for a specific purified diphosphoryl lipid A (thinlayer chromatography-3 fraction) but negative for the purified monophosphoryl lipid A (also a thin-layer chromatography-3 fraction). These results show that the LAL test is not a valid measure of all parameters of toxicity of a lipid A or lipid Alike compound and can yield false-positive results. However, these findings are not in conflict with the widespread use of the LAL assay for pyrogens in the pharmaceutical industry since a good correlation exists between LAL results and pyrogenicity when undegraded endotoxin is evaluated in parallel assays.

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Rapid Detection of Gram-Negative Bacteriuria by Use of the Limulus Endotoxin Assay

Cited by 57 publications

References 9 publications

The effect of rinsing with Listerine antiseptic on the properties of developing dental plaque

The effect of rinsing with Listerine antiseptic on the properties of developing dental plaque

Preliminary characterization of material eluted from the roots of periodontally diseased teeth

Influence of fine structure of lipid A on Limulus amebocyte lysate clotting and toxic activities

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