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  • Title: A quantitative in vitro assay for detecting biological activity of endotoxin using rabbit peripheral blood.
    Author: Ochiai M, Kataoka M, Toyoizumi H, Yamamoto A, Horiuchi Y.
    Journal: Jpn J Infect Dis; 2001 Aug; 54(4):131-6. PubMed ID: 11684780.
    Abstract:
    The pyrogen test or the endotoxin test has been playing a crucial role in detecting endotoxin in parenteral drugs. The current test methods, however, have disadvantages such as requiring a relatively high number of animals or an inadequacy in direct evaluation of in vivo activity. We made an attempt to establish a new in vitro assay method that can overcome the shortcomings of the current assay methods. We standardized the system of tumor necrosis factor-alpha (TNF-alpha) induction from the peripheral blood of rabbits for assaying endotoxin activity. This in vitro assay showed a linear dose-response regression between 0.1 and 5.0 endotoxin units per milliliter of endotoxin and a definite homogeneity of variance by logarithmically transforming the endotoxin and TNF-alpha concentrations in the reaction mixtures at 5 h of incubation at 37 degrees C. The assay showed a definite correlation with the pyrogen test but not with the endotoxin test when endotoxins from various bacteria were tested.
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