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The Visual Colorimetric Detection of Multi-nucleotide Polymorphisms on a Pneumatic Droplet Manipulation Platform
Published on: September 27, 2016
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On-chip detection of a single nucleotide polymorphism without polymerase amplification.
Jinhee Han1, Matthew Tan2, Lakshmana Sudheendra1
1Department of Mechanical and Aerospace Engineering, University of California, Davis, California 95616, USA.
Nano Research
|January 13, 2015
Summary
A novel photonic crystal (PC) array enhances fluorescent signals for sensitive single nucleotide polymorphism (SNP) detection. This PC/DNA assay reliably identifies genetic mutations in feline models, showing promise for portable nucleic acid analysis.
Area of Science:
- Nanotechnology
- Biomedical Engineering
- Genetics
Background:
- Single nucleotide polymorphism (SNP) detection is crucial for diagnosing genetic diseases.
- Existing methods for SNP detection can be complex and expensive.
- Photonic crystals (PCs) offer potential for enhanced optical detection systems.
Purpose of the Study:
- To develop and validate a nanoparticle-assembled photonic crystal (PC) array for sensitive SNP detection.
- To assess the PC/DNA assay's ability to identify specific genetic mutations in a feline model.
- To compare the PC/DNA assay's performance with established methods like quantitative polymerase chain reaction (PCR).
Main Methods:
- Assembled nanoparticle-based PC arrays with nanowells.
- Utilized an electrophoretic particle entrapment system to load probe DNA-hybridized nanoparticles.
- Employed the PC/DNA assay to detect SNPs in synthesized nucleotide sequences and feline genomic DNA (gDNA).
Main Results:
- The PC/DNA assay demonstrated high sensitivity (0.9 fg/mL) and detected a 1 base pair difference in synthesized sequences.
- Successfully identified SNPs in gDNA from urine and blood samples of cats with and without polycystic kidney disease (PKD).
- Achieved a limit of detection (LOD) of 0.005 ng/mL for gDNA in real samples, comparable to real-time PCR.
Conclusions:
- The nanoparticle-assembled PC array is a highly sensitive and reliable platform for SNP detection.
- The PC/DNA assay effectively identified PKD-associated mutations in feline samples.
- PC technology holds potential for developing simple, cost-effective, and portable nucleic acid analysis tools.
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