[Use of beta-lactamase from Bacillus licheniformis 749/C for solid-phase immunoenzyme analysis]

Area of Science:

• Biotechnology
• Immunology
• Microbiology

Background:

• Enzyme-linked immunosorbent assay (ELISA) is a common diagnostic tool.
• Sensitive detection of the plague pathogen is crucial for public health.
• Beta-lactamase offers a detectable enzymatic marker.

Purpose of the Study:

• To develop a highly sensitive ELISA for detecting the plague causative agent.
• To utilize beta-lactamase from Bacillus licheniformis 749/c as a marker in ELISA.
• To create conjugates of beta-lactamase with plague antigen and antibodies.

Main Methods:

• Preparation of beta-lactamase conjugates with plague capsule antigen and monoclonal antibodies using glutaric aldehyde.
• Enzyme-linked immunosorbent assay (ELISA) for antigen detection.
• Microiodometric determination of beta-lactamase enzymatic activity for result visualization.

Main Results:

• Conjugates retained high immunospecific and beta-lactamase activity.
• The modified ELISA demonstrated high sensitivity, detecting as low as 8 ng/ml of plague capsule antigen.
• Microiodometric assay provided visual recording of ELISA results.

Conclusions:

• The developed ELISA method is highly sensitive and specific for detecting the plague pathogen.
• Beta-lactamase serves as an effective marker for sensitive and visually recordable plague diagnostics.
• This approach offers a promising tool for rapid plague detection.