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Ultrasensitive ELISA Developed for Diagnosis
Kanako Iha1, Mikio Inada1, Naoki Kawada1
1Department of Biology, Waseda University, Tokyo 162-8480, Japan.
Diagnostics (Basel, Switzerland)
|July 21, 2019
Summary
This study introduces a new ultrasensitive enzyme-linked immunosorbent assay (ELISA) for detecting trace disease biomarkers. The novel method achieves high sensitivity for proteins and nucleic acids, enabling rapid and accurate diagnostics.
Area of Science:
- Biotechnology
- Assay Development
- Medical Diagnostics
Background:
- Quantitative detection of trace biomarkers is crucial for disease diagnosis.
- Existing methods often lack sensitivity or require specialized equipment.
Purpose of the Study:
- To develop a simple, rapid, and ultrasensitive detection method for disease biomarkers.
- To demonstrate the versatility of the assay for both protein and nucleic acid detection.
Main Methods:
- A novel ultrasensitive assay combining sandwich enzyme-linked immunosorbent assay (ELISA) with thionicotinamide-adenine dinucleotide (thio-NAD) cycling.
- Measurement of accumulated thio-NADH at 405 nm for signal detection.
- Adaptation of the assay for nucleic acid detection using hybridization.
Main Results:
- Achieved a limit of detection of approximately 10-18 moles/assay for target proteins.
- Successfully detected HIV-1 p24 and adiponectin (including in urine), with limits of detection of 10-18 and 2.3 × 10-19 moles/assay, respectively.
- Demonstrated applicability to nucleic acid detection without amplification.
Conclusions:
- The developed ultrasensitive ELISA offers a versatile, simple, and rapid diagnostic tool.
- The assay's ability to detect trace biomarkers in patient samples facilitates clinical diagnosis.
- This de novo method enables accurate detection of various disease markers.
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