Quantitative Limulus lysate assay for endotoxin and the effect of plasma.

Hurley, James C.; Tosolini, F. A.; Louis, William J.

doi:10.1136/jcp.44.10.849

Cited by 46 publications

(41 citation statements)

References 13 publications

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“…Most importantly, quantitative analysis by HPLC/MS/MS revealed that maximal plasma levels of LPS were actually measured after 6 h, thus indicating that 1 ) the bulk of circulating LPS was still present in the plasma compartment at this time, and 2 ) in accordance with earlier reports ( 8,25 ), the LAL assay can produce false negative results in native plasma. These fi ndings are in agreement with earlier studies, which reported that in vitro incubation of plasma can mask LPS detection in a concentration-dependent manner (26)(27)(28). The combination of the LAL and HPLC/ MS/MS analyses in the present study provided new evidence of the intrinsic capacity of plasma to neutralize the activity of LPS.…”

Section: Discussionsupporting

confidence: 82%

Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay

Barros

Gautier

Sali

et al. 2015

Journal of Lipid Research

109

103

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Systemic infl ammatory response syndrome (SIRS) is defi ned by a cluster of clinical signs that include tachycardia, leukocytosis, tachypnoea, and pyrexia ( 1 ). Although SIRS is a major consequence of sepsis (i.e., a deleterious, nonresolving infl ammatory response to infection that can lead to multiple organ dysfunction and septic shock), it can be caused by many pathological events that are not associated with the bacterial infection ( 1 ). It is of major importance to distinguish between noninfectious SIRS, sepsis, and septic shock in critically ill patients because these groups of patients are markedly different in terms of care and clinical outcome. Positive microbiological identifi cation tests [using conventional microbiological tests or bacterial DNA detection ( 2 ) as well as a number of infl ammatory and infectious biomarkers, including cytokines, procalcitonin, immune cell markers or a combination of these ( 3 )] have been proposed, but at this stage, none has proved to be reliable enough to ascertain the occurrence and extent of infection in high-risk patients with SIRS. Interestingly, and as documented further by our group in the prospective multicenter cohort EPISS study ( 4 ), Gramnegative bacilli are the most frequently identifi ed pathogens in patients with septic shock. In this context, the direct quantitation of the culprit component of the Gram-negative bacterium [i.e., lipopolysaccharide (LPS), which triggers SIRS by interacting with the CD14/Toll-like receptor 4 /myeloid differentiation factor 2 receptor complex at the surface of leukocytes ( 1 ) Abbreviations: 3HM, 3-hydroxymyristic acid or 3-hydroxymyristate; LAL, limulus amebocyte lysate; LOD, limit of detection; LOQ, limit of quantifi cation; LPS, lipopolysaccharide; PLTP, phospholipid transfer protein; SIRS, systemic infl ammatory response syndrome; SOFA, sequential organ failure assessment .

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Section: Discussionsupporting

confidence: 82%

Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay

Barros

Gautier

Sali

et al. 2015

Journal of Lipid Research

109

103

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“…Drugs that interfere with the clotting system, i.e., through inhibition (binding of divalent cations such as ethylenediaminetetraacetic acid, citrate, protease inhibitors) or enhancement (high protein content, proteases), cannot be tested with the lAl test (Cooper et al, 1997;Duner, 1995). Furthermore, a number of endotoxin-binding components from plasma are known to mask lPS in the lAl test (Hurley et al, 1991), and due to such interference with the test system, many drugs have to be diluted for testing (al-Khalifa et al, 1989;elin and Wolff, 1973). The LAL cascade also is triggered by (1,3)-β-dglucan Cooper et al, 1997) and polysaccharides, for example from cellulose filter materials, which can result in false-positive signals (Ikemura et al, 1989;Anderson et al, 2002).…”

Section: In Mononuclear Cells When the 4/2 Acyl Chain Pattern Of Entementioning

confidence: 99%

Evidence for the detection of non-endotoxin pyrogens by the whole blood monocyte activation test

Hasiwa

Daneshian

Bruegger

et al. 2013

ALTEX

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“…Clarifications of published data were sought from the original authors. This search was in addition to publications obtained from a library of several hundred publications related to clinical aspects of endotoxemia that I accumulated from repeatedly searching the literature over two decades (29,33).Study selection and classification of bacteria. The following inclusion criteria were used: (i) comparison of the Limulus assay with blood cultures from patients with suspected GN bacteremia, (ii) the sensitivity of the Limulus assay to an internal endotoxin standard stated, and (iii) a minimum of two patients with GN bacteremia.…”

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confidence: 99%

Diagnosis of Endotoxemia with Gram-Negative Bacteremia Is Bacterial Species Dependent: a Meta-Analysis of Clinical Studies

Hurley

2009

J Clin Microbiol

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Endotoxemia is undetectable for up to 60% of cases of bacteremia caused by gram-negative (GN) species, a discordance attributed to the limitations of the Limulus assay for endotoxemia. The lipid A structure of the endotoxin molecule is critical for the sensing of GN bacteria by the host immune system although not so for sensing by the Limulus assay. The lipid A structure of commensal Enterobacteriaceae is hexa-acyl, whereas non-Enterobacteriaceae have a broader range of structures. By using a previously published classification of lipid A structures (R. . This finding is consistent across all the unrestricted studies, even including studies with seemingly contrary results for endotoxemia diagnosis among cases of bacteremia caused by GN bacteria overall. Surprisingly, with bacteremia caused by commensal Enterobacteriaceae, the diagnosis of endotoxemia appears to be unrelated to the Limulus assay sensitivity. Across these 45 studies, the association of endotoxemia with GN bacteremia is variable but consistent for different types of GN bacteremia.There are key structural differences between the lipid A components of the endotoxin molecule (lipopolysaccharide) of different gram-negative (GN) bacteria. Members of the Enterobacteriaceae characteristically have a lipid A structure with a hexa-acyl structure, whereas other lipid A structures are present for non-Enterobacteriaceae (45). These differences in lipid A structure are now known to be critically important for the recognition of GN bacteremia by the host immune system (45) by the MD-2-Toll-like receptor 4 receptor but not for sensing by the clotting proteins of the blood cells of the Limulus polyphemus horseshoe crab, from which the Limulus amebocyte lysate assay is derived (56). If the recognition of hexaacyl lipid A by the host immune system has a role in the pathogenesis of bacteremia, it might be expected that the proportion with detectable endotoxemia among patients with GN bacteremia might depend on the lipid A structure of the isolate.Attempts to define the concordance between GN bacteremia and endotoxemia in patients with sepsis have been elusive. Indeed, two of those studies (15, 54), with over a hundred patients each, concluded that there is no concordance. The purpose of this review is to attempt to reconcile the disparate findings from studies of clinically detected sepsis by examining the proportion with endotoxemia detected by using the Limulus assay for patients with different types of GN bacteremia for which the lipid A structures are known. Of particular interest is the proportion with endotoxemia among cases of bacteremia caused by commensal Enterobacteriaceae versus non-Enterobacteriaceae. The statistical techniques of meta-analysis are used to derive study-specific and summary estimates of these proportions expressed as an OR and more so to obtain estimates of the consistency in these ORs across the panel of studies (24). MATERIALS AND METHODS Data sources.A comprehensive search of the literature from 1966 to April 2009 was performed by using...

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Quantitative Limulus lysate assay for endotoxin and the effect of plasma.

Abstract: The effects of plasma and chromogenic substrate on the kinetics of the endotoxin-activated Limulus

Cited by 46 publications

References 13 publications

Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay

Quantitative lipopolysaccharide analysis using HPLC/MS/MS and its combination with the limulus amebocyte lysate assay

Evidence for the detection of non-endotoxin pyrogens by the whole blood monocyte activation test

Diagnosis of Endotoxemia with Gram-Negative Bacteremia Is Bacterial Species Dependent: a Meta-Analysis of Clinical Studies

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