Rapid homogenous time-resolved fluorescence (HTRF) immunoassay for anthrax detection

Area of Science:

• Biomedical diagnostics
• Immunotechnology
• Infectious disease detection

Background:

• Anthrax, caused by Bacillus anthracis, is a severe disease requiring early diagnosis for effective treatment.
• Current diagnostic methods may lack the speed and sensitivity needed for timely intervention.

Purpose of the Study:

• To develop rapid and sensitive homogenous time-resolved fluorescence (HTRF) immunoassays for detecting Bacillus anthracis.
• To enable early diagnosis of anthrax by detecting key biomarkers like Protective Antigen (PA) toxin and bacterial spores.

Main Methods:

• Development of HTRF immunoassays utilizing europium cryptate (EuK) donor and AlexaFluor647 acceptor energy transfer.
• Utilized monoclonal antibodies for PA toxin detection and polyclonal antibodies for spore detection.
• Assayed samples from infected rabbits and tested specificity against Bacillus cereus.

Main Results:

• HTRF assay detected 2 ng/ml of PA toxin in rabbit serum within 15 minutes.
• HTRF assay detected 2 × 10^6 spores/ml within 30 minutes.
• Assays demonstrated high sensitivity, specificity, and rapid detection capabilities.

Conclusions:

• HTRF immunoassays provide a rapid, sensitive, and simple method for detecting both single-epitope (proteins) and multi-epitope (particles) targets.
• These assays are valuable for timely medical care in suspected anthrax cases.
• The developed HTRF technology offers a promising approach for infectious disease diagnostics.