RAPID DETECTION OF BACILLUS ANTHRACIS SPORES BY IMMUNOCHROMATOGRAPHY METHOD

Abstract

    Anthrax in human and animal caused by Bacillus anthracis . It is a dangerous disease. B.Anthracis spores may be using in bioterrorism and bioweapons. Therefore, there is an urgentneed for convenient, sensitive, and specific methods to detect the spores of B. Anthracis. Thisarticle reports an immunochromatography device based on the principle of sandwich assay(lateral flow test). This test were developed by using monoclonal anti - B. Anthracis sporesSA26 and SA27. The spores captured by a monoclonal anti-spore antibody gold conjugated(SA27) in the conjugate pad. This complex will was captured by other monoclonal anti -spore (SA26) coated on the nitrocellulose membrane (sanwhich assay), so T line appear . Thesolution migrates by capillary action through the test strip. This complex was captured bygoat anti - mouse IgG antibody, C line appear. If it was negative (without spores in sample),the T line disappeared. If it was positive (with spores), the T line developed in the T region.This method had high sensitivity and specificity. Result was read rapidly. We chose severalappropriate conditions as: concentration of capture reagents on test line and control line,concentration of colloidal conjugated gold on the conjugate pad, treated sample pad, sampleextract buffer to make a rapid test. We successfully created a rapid test to detect Bacillusanthracis spores. The limit of detection of spores is 105 spores/ml. Reading results are set at15 minutes. Sensitivity is 100% and specificity is 97%. This device is simple and activites inthe field. Store the test below room temperature.

Keywords: Capture reagents, Bacillus anthracis, rapid test, spores, lateral flow test.