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Limulus amebocyte lysate test for endotoxemia

Investigations with a femtogram sensitive spectrophotometric assay

Limulus Amebozyten Lysat Test für Endotoxinaemie

Untersuchungen mit einem Fematogramm-sensitiven spektrophotometrischen Test

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Summary

The question of specificity of Limulus amebocyte lysate (LAL) test in the diagnosis of endotoxemia has been a limiting factor of its clinical application. Using a femtogram-sensitive spectrophotometric LAL assay 35 of 36 septic postoperative patients showed an excellent correlation (almost 100%) between positive LAL tests and culture-proven gramnegative bacteremia. Twenty patients of this group demonstrated a significant correlation (2p<0.05) between elevated total white blood cell counts and Escherichia coli Endotoxin equivalents (EcEe).

All 22 liver cirrhotic patients with potentially enteric endotoxemia yielded LAL positive reactions (100%) but 9 of 22 correlated significantly as to leucocytosis and elevated EcEe (2p<0.01). A significant correlation between EcEe and leucocyte counts was found for all neonates and all parturients respectively (2p<0.001, 2p<0.01). In vitro tests showed that leucocytes gave positive LAL tests. Supernates of a Ficoll sedimented crude leucocyte preparations, induced LAL positivity in a dose dependent manner, while all reagents per se used in the preparation, were negative. This study suggests that heat-labile factors from leucocytes apart from endotoxin are responsible for positive LAL reactions.

Zusammenfassung

Die fragliche Spezifität des Limulus Amebozyten Lysat (LAL) Testes bei der Diagnose der Endotoxinaemie stellt einen limitierenden Faktor für seinen klinischen Einsatz dar. Mit Hilfe eines Femtogramm-sensitiven spektrophotometrischen LAL Testes zeigten 35 von 36 septischen postoperativen Patienten eine gute Korrelation (fast 100%) zwischen positiven LAL Testen und Blutkultur positiven gram-negativen Bakteriaemien. 20 Patienten dieser Gruppe zeigten eine signifikante Korrelation (2p<0.05) zwischen erhöhten Leukozytenzahlen und Escherichia coli Endotoxin Äquivalenten (EcEä). Bei 22 Patienten mit Leberzirrhose und einer potentiell enterogenen Endotoxinaemie konnten LAL positive Reaktionen (100%) nachgewiesen werden, aber 9 von 22 korrelierten signifikant (2p<0.01) zwischen Leukozytose und erhöhten EcEä. Eine signifikante Korrelation zwischen EcEä und Leukozytenzahlen wurde für alle Neonaten und Schwangeren (2p<0.001, 2p<0.01) festgestellt. In vitro Tests zeigten, daß Leukozyten positive LAL Tests bewirkten. Überstände der mit Ficoll sedimentierten groben Leukozyten-Präparationen induzierten eine LAL Positivität in einer dosisabhängigen Form, während alle Reagentien per se, die für die Präparation gebraucht wurden, negativ waren. Diese Arbeit legt die Vermutung nahe, daß hitze-labile Faktoren von Leukozyten neben Endotoxin für positive LAL Reaktionen verantwortlich sind.

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Authors and Affiliations

  1. Department of Surgery, Medical School, Hannover

    L. Lehr

  2. Department of Immunology and Serology, Institute of Hygiene and Medical Microbiology, Klinikum Mannheim, University of Heidelberg, Mannheim

    R. M. Urbaschek

  3. Institut für Klinische Chemie, Medizinische Hochschule Hannover, Karl-Wiechert-Allee 9, D-3000, Hannover 61, Germany

    P. C. Fink & J. Kozak

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  1. P. C. Fink
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  2. L. Lehr
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  3. R. M. Urbaschek
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  4. J. Kozak
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Fink, P.C., Lehr, L., Urbaschek, R.M. et al. Limulus amebocyte lysate test for endotoxemia. Klin Wochenschr 59, 213–218 (1981). https://doi.org/10.1007/BF01476578

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  • DOI: https://doi.org/10.1007/BF01476578

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