METHOD OF DETECTING PRESENCE OF ANTHRAX INFECTION IN BIOLOGICAL FLUIDS AND ENVIRONMENT Russian patent published in 2011 - IPC C12Q1/42 G01N33/543 G01N33/569 

FIELD: medicine.

SUBSTANCE: method of detecting anthrax pathogen in environment and biological fluids is based on functional detection of activity of one of anthrax toxin components, weak protease of anthrax lethal factor (LF). Applied principle of highly sensitive detection of LF proteolytic activity is based on the system of amplification of sugnal, originating from the act of substrate cutting, by means of present in substrate composition auxiliary enzyme of alkaline phosphatase of Escherichia coli (AP). In composition of recombinant LF substrate in addition to alkaline phosphotase and specific for LF substrate sequence RRKKVYPYPME, there is a peptide which is biotinilated in vivo and in vitro by means of biotin-ligase of E.coli and ensuring substrate immobilisation on solid phase (plaque surface) due to interaction with avidin and its derivatives, and sequence of six histidines for purification of recombinant substrate from periplasm of E.coli by means of metal-chelate resin.

EFFECT: method in accordance with the invention possesses high sensitivity, with application of fluorescein diphosphate makes it possible to detect up to 1 pM of lethal factor, rapidity of execution, high specificity, low risk of false-positive signals.

1 dwg, 3 ex