Endotoxin detection in end-stage kidney disease.
OBJECTIVE
Endotoxin detection assays are not validated for use in end-stage kidney disease (ESKD). We investigated the accuracy and precision of the kinetic turbidimetric Limulus amoebocyte lysate (LAL) assay to detect endotoxin in plasma from patients with ESKD. Optimisation of endotoxin recovery from plasma using the detergent Tween 80 was also explored.


METHODS
Plasma samples from 7 patients with ESKD and 7 healthy subjects were spiked with different concentrations of endotoxin. Repeated measurements for endotoxin at each level of spike were performed to assess the accuracy and precision of spike recovery. Endotoxin recovery in plasma samples diluted in Tween 80 and water was compared.


RESULTS
Mean endotoxin spike recovery was 111.6% and 125.2% in ESKD and healthy subjects, respectively. There was no statistical difference in spike recovery between ESKD and healthy plasma. Precision of the LAL assay in plasma spiked with low (0.05 EU/mL) and high (0.5 EU/mL) concentration of endotoxin spikes was 24.1% and 8.9%, respectively. The use of Tween 80 as a diluent for plasma significantly improved spike recovery in ESKD plasma (100.1% vs 70.4%, p<0.001).


CONCLUSIONS
The kinetic LAL turbidimetric assay is a valid tool for the detection of blood endotoxin in patients with ESKD, although in blood specimens with low-level endotoxemia (≤0.05 EU/mL) the assay may be less accurate and precise. Tween 80 can be used as a diluent to optimise recovery of endotoxin in ESKD plasma.
Department of Renal Medicine, Lister Hospital, Hertfordshire, UK Department of Postgraduate Medicine, University of Hertfordshire, Hertfordshire, UK.
Jonathan
J
Wong
Quality Control Department, Lister Hospital, Hertfordshire, UK.
Hassan
H
Jeraj
Department of Renal Medicine, Lister Hospital, Hertfordshire, UK Department of Postgraduate Medicine, University of Hertfordshire, Hertfordshire, UK.
Enric
E
Vilar
Department of Biochemistry, Cambridge University Hospitals, Hertfordshire, UK Department of Postgraduate Medicine, University of Bedfordshire, Bedfordshire, UK.
Adie
A
Viljoen
Department of Renal Medicine, Lister Hospital, Hertfordshire, UK Department of Postgraduate Medicine, University of Hertfordshire, Hertfordshire, UK.
Ken
K
Farrington
D001774:Blood Chemical Analysis / Q000379:methods*; D004731:Endotoxins / Q000097:blood*; D005260:Female; D006801:Humans; D007676:Kidney Failure, Chronic / Q000097:blood*; D007700:Kinetics; D008297:Male; D008875:Middle Aged; D009391:Nephelometry and Turbidimetry
D004731:Endotoxins
ENDOTOXIN; NEPHROLOGY; RENAL