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Multiplex PCR and Reverse Line Blot Hybridization Assay mPCR/RLB

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Published on: August 6, 2011

Paper-Based Multiplex Surface-Enhanced Raman Scattering Detection Using Polymerase Chain Reaction Probe Codification.

Eun Ju Kim^1,2, Hanbi Kim^1,2, Eunkyoung Park^1,2

¹Biomedical Engineering Research Center, Smart Healthcare Research Institute, Samsung Medical Center, 81 Irwon-ro, Gangnam-gu, Seoul 06351, Republic of Korea.

Analytical Chemistry

|February 19, 2021

Summary

This study introduces a novel plasmonic paper platform for highly sensitive, multiplexed detection of fluorescent dyes using surface-enhanced Raman scattering (SERS). This cost-effective method enables simultaneous pathogen identification via magnetic bead separation and RT-PCR.

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Area of Science:

Nanotechnology
Analytical Chemistry
Biotechnology

Background:

Surface-enhanced Raman scattering (SERS) offers high sensitivity for molecular detection.
Plasmonic nanomaterials on paper substrates present opportunities for portable biosensing.
Multiplexed detection is crucial for identifying multiple analytes simultaneously.

Purpose of the Study:

To develop a multiplex SERS platform for sensitive detection of multiple fluorescent dyes.
To utilize a plasmonic paper substrate and a double-labeled probe system.
To enable simultaneous detection of multiple targets using a single-wavelength light source.

Main Methods:

Fabrication of a plasmonic paper substrate with silver nanodots on cellulose fibers.
Design of double-labeled probes with fluorescent dyes and biotin.
Employing streptavidin-coated magnetic beads (SMBs) for separation and reverse transcription polymerase chain reaction (RT-PCR) for amplification and detection.

Main Results:

Achieved highly sensitive SERS detection of fluorescent dyes from picomolar to millimolar concentrations.
Demonstrated multiplex detection capability using the plasmonic paper and magnetic bead separation.
Validated the method's efficiency and economy for simultaneous detection of multiple pathogens.

Conclusions:

The developed plasmonic paper SERS platform enables sensitive and multiplexed detection of fluorescent dyes.
The combination of magnetic bead separation and SERS is effective for simultaneous pathogen identification.
This approach offers a cost-efficient and adaptable solution for multiplexed biosensing applications.