Method for monitoring periodontal disease by monitoring endotoxins and inflammatory agents

A method of quantitatively determining the amount of bacterial endotoxins in a periodontal pocket of a subject which comprises:

• (a) collecting a sample from the periodontal pocket of the subject on a filter paper strip by placing the strip momentarily in the periodontal pocket;
• (b) dry heating the strip containing the sample so as to inactivate endogenous enzymes which would interfere with Limulus amebocyte lysate-endotoxin interaction;
• (c) contacting the dry heated strip containing the sample with a Limulus amebocyte lysate under conditions so as to activate a proenzyme contained in the Limulus amebocyte lysate, the resulting enzyme being capable of cleaving a bond between an arginyl group and a nitrogen-containing group, such that the sample contains the enzyme;
• (d) contacting the enzyme contained in the resulting sample with a substrate selected from the group consisting of benzoylarginine-.beta.-naphthylamide, benzoylarginine-p-nitroanilide, or acetyl-Ileu-Glu-Gly-Arg-(p-nitroanilide) so as to form an amine;
• (e) treating the resulting amine to produce a visually detectable product;
• and
• (f) quantitatively determining the amount of the product formed and thereby the amount of bacterial endotoxins present in the periodontal pocket.