Detection and clinical relevance of human endotoxemia.
An optimized chromogenic assay for the detection of endotoxins in human blood is described. The assay comprises the removal of the inhibitory activity of plasma components by a dilution plus heating procedure, endotoxin-dependent activation of Limulus amebocyte lysate, and chromogenic measurement of the activated lysate. The assay has a detection limit of 3 ng endotoxin/L plasma. At the 10 ng/L level within-assay CV and between-assay CV of 14 and 21% were obtained respectively. In a prospective clinical trial including 473 consecutive febrile patients the assay has previously been demonstrated to have positive and negative predictive values of 48% and 99% for impending Gram-negative sepsis, respectively. In a similar study in 76 consecutive patients with Gram-negative infection of the urinary tract, these values were 73% and 95%, respectively. We conclude that this assay may provide the means to select those patients who are most likely to benefit from anti-endotoxin treatment. To facilitate endotoxin testing in other laboratories, a preliminary evaluation of a commercial endotoxin assay versus our own method was performed with 108 duplicate blood samples obtained from septic patients. With this assay detection limits of 2-3 ng endotoxin/L plasma could be obtained, as well as a good correlation (r = 0.94) and level of consensus to establish endotoxemia (93%) as compared to the house method. The commercial assay may therefore facilitate the introduction of endotoxin testing in other laboratories.
Sturk
A
A
Department of Clinical Chemistry, Academic Medical Center, Amsterdam, The Netherlands.
van Deventer
S J
SJ
Wortel
C H
CH
Levels
J H
JH
ten Cate
J W
JW
Büller
H R
HR
Sanders
G T
GT
D001424:Bacterial Infections / Q000175:diagnosis*; D002863:Chromogenic Compounds; D004731:Endotoxins / Q000097:blood*; D006090:Gram-Negative Bacteria; D006801:Humans; D008070:Lipopolysaccharides / Q000097:blood*; D011237:Predictive Value of Tests; D011446:Prospective Studies; D014115:Toxemia / Q000175:diagnosis*; D014552:Urinary Tract Infections / Q000175:diagnosis*
D002863:Chromogenic Compounds; D004731:Endotoxins; D008070:Lipopolysaccharides