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Author Spotlight: Advancing Pathogen Diagnostics with Standardized LAMP

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Nanoplasmonic microarray-based solid-phase amplification for highly sensitive and multiplexed molecular diagnostics:

Ji Young Lee¹, Hyowon Jang², Sunjoo Kim³

¹Advanced Bio and Healthcare Materials Research Division, Korea Institute of Materials Science (KIMS), 797, Changwon-Daero, Seongsan-Gu, Changwon-Si, Gyeongsangnam-Do, 51508, Republic of Korea.

Mikrochimica Acta

|October 30, 2024

Summary

This summary is machine-generated.

This study introduces a novel nanoplasmonic microarray method for sensitive gene detection. The approach achieves high accuracy and speed for identifying SARS-CoV-2 genes, enabling rapid on-site diagnostics.

Keywords:

Differentiation of mutations Multiplex molecular diagnostics Nanoplasmonic microarrays SARS-CoV-2 Solid-phase amplification

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Area of Science:

Nanotechnology and Molecular Diagnostics
Biotechnology and Genetic Engineering

Background:

Current molecular diagnostic methods often face limitations in sensitivity, speed, and multiplexing capabilities.
Solid-phase amplification techniques require optimization to overcome sensitivity issues for widespread application.
The need for rapid, accurate, and on-site diagnostic tools, especially for infectious diseases like SARS-CoV-2, is critical.

Purpose of the Study:

To develop and evaluate a novel nanoplasmonic microarray-based solid-phase recombinase polymerase amplification (RPA) system.
To enhance sensitivity and multiplexing capabilities for gene detection using nanoplasmonic substrates.
To assess the performance of this system for the rapid and accurate detection of SARS-CoV-2 genes, including Omicron variants.

Main Methods:

Development of nanoplasmonic microarrays via one-step immobilization of streptavidin/biotin primers.
Optimization of amplicon size for enhanced plasmon-enhanced fluorescence (PEF) on the nanoplasmonic substrate.
Solid-phase RPA assay for detecting E, N, and RdRP genes of SARS-CoV-2, with stringent washing and primer-dimer minimization.

Main Results:

Achieved high sensitivity with a limit of detection of four copies per reaction within 30 minutes.
Demonstrated high specificity by minimizing primer-dimer formation and employing a stringent washing process.
Clinical testing with 30 nasopharyngeal swab samples showed 100% consistency with PCR results, including differentiation of Omicron BA.1 and BA.2 mutations.

Conclusions:

The nanoplasmonic microarray-based solid-phase RPA approach overcomes sensitivity limitations of traditional solid-phase amplification.
This method offers a rapid, highly sensitive, and multiplexing-capable solution for gene detection.
The system's simplicity and isothermal nature make it a valuable tool for on-site molecular diagnostics.