Detection of pathogen-specific antibodies by loop-mediated isothermal amplification

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Immunology

Background:

• Loop-mediated isothermal amplification (LAMP) is a sensitive DNA amplification technique suitable for point-of-care (POC) diagnostics.
• Measuring protein analytes, particularly IgG antibodies, is crucial for diagnosing infectious diseases.
• Existing methods may lack the sensitivity or speed required for rapid POC diagnosis.

Purpose of the Study:

• To adapt LAMP for the detection and quantification of protein analytes.
• To develop a novel protein-DNA fusion construct, termed "LAMPole," for enhanced protein detection.
• To demonstrate the utility of LAMPoles for sensitive serological response measurement.

Main Methods:

• Creation of LAMPoles by fusing DNA elements to IgG-binding polypeptides (Protein L/G domain).
• Utilizing LAMP for the amplification of LAMPoles in a real-time turbidity assay.
• Quantifying serological responses to a protozoan parasite in serum samples.

Main Results:

• Demonstrated a >6-log fold difference in signal between vaccinated and control mice sera.
• Observed significant signal differences in clinical patient samples compared to controls.
• Validated the sensitivity and specificity of the LAMPole assay for protein quantification.

Conclusions:

• LAMPoles represent a promising platform for sensitive protein analyte measurement.
• This technology can significantly improve acute infection diagnosis in both clinical laboratory and field settings.
• The developed method enhances POC diagnostic capabilities for various infectious diseases.