Development of oligonucleotide lateral-flow immunoassay for multi-parameter detection

Area of Science:

• Biotechnology
• Immunodiagnostics
• Molecular Biology

Background:

• Current diagnostic methods often lack the capacity for simultaneous detection of multiple analytes.
• There is a need for rapid, sensitive, and multiplexed assays in clinical and research settings.

Purpose of the Study:

• To develop and validate a highly sensitive and rapid oligonucleotide lateral-flow immunoassay (OLFIA) for simultaneous detection of multiple antigens and/or antibodies.
• To demonstrate the capability of the OLFIA system to detect more than two types of targets in a single assay device.

Main Methods:

• Development of an OLFIA system utilizing colloidal gold as an indicator.
• Incorporation of colloidal gold-labeled antibodies and oligonucleotide-labeled antibodies.
• Immobilization of complementary oligonucleotide-labeled proteins on a nitrocellulose membrane.
• Utilizing DNA-DNA interactions for solid-phase binding of target complexes.

Main Results:

• The OLFIA system demonstrated high sensitivity and rapid detection capabilities.
• Simultaneous detection of multiple antigens and/or antibodies was achieved in a single assay.
• The assay design allows for multiplexed detection through strategic immobilization of different labeled components.

Conclusions:

• The developed OLFIA is a sensitive and rapid platform for multiplexed detection of antigens and/or antibodies.
• This technology offers a significant advancement for simultaneous diagnostic testing in various biological specimens.
• The DNA-DNA interaction-based capture mechanism provides a robust foundation for multi-analyte lateral-flow assays.