Automation of the Limulus amoebocyte lysate test by using the Abbott MS-2 microbiology system

Abstract

A rapid, automated method for the performance of the Limulus amoebocyte lysate endotoxin assay has been developed by using the Abbott MS-2 Microbiology System. This instrument automatically determines sequential changes in the optical density of up to 176 samples at 1- or 5-min increments during a 1-h assay period. Graphic representation of optical density changes can be viewed on a cathode-ray tube or reproduced by using a hard-copy printer. Limulus amoebocyte lysate preparations that were obtained from different commercial producers and that had similar endotoxin sensitivities by the conventional gelation method varied somewhat in reactivity when determinations were based upon rate changes in optical density. Lysates from Associates of Cape Cod, Difco Laboratories, and M. A. Bioproducts were the most readily adaptable to the MS-2 System. Use of the MS-2 system increased the sensitivity of these preparations from 60- to 250-fold, and as little as 1 pg/ml was detected. Adaptation of the MS-2 instrument for this purpose provides an objective, reproducible, automated method for the performance of Limulus amoebocyte lysate tests on a variety of fluids.