Development and Testing of Monoclonal Antibodies-Based Diagnostic Preparation for <i>Bacillus anthracis</i> Spores Detection Using Latex Agglutination Method

Abstract

Immobilization of anti-B. anthracis monoclonal antibodies (MAbs) on latex microparticles was studied, the optimal load of these MAbs was determined to be 20 µg for 50 µl of the stock latex suspension. The highest sensitivity of latex agglutination test was observed for 1E6 MAbs. Latex suspensions with immobilized MAbs were lyophilized. Their sensitivity and specificity were shown to be highly competitive with those of the stock liquid latex suspension. Latex diagnosticum for Bacillus anthracis spores detection was constructed on the basis of these lyophilized reagents, developed and approved was the regulatory documentation that included their application instructions and technical specifications. Carried out were inter-laboratory and commission tests of experimental prototypes of the designed latex diagnosticum. These preparations demonstrated high sensitivity (from 1·105 to 2·106 spores/ml and even more) and specificity (absence of cross-reactions with spores of different species of sporogenous bacilli at concentration of 108 spores/ml).

Keywords

• monoclonal antibodies (mabs)
• latex microparticles
• latex agglutination test
• sensitivity
• specificity
• commission tests