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Direct Detection of Low Abundance Genes of Single Point Mutation
- Source :
- Nano Letters. 21:9061-9068
- Publication Year :
- 2021
- Publisher :
- American Chemical Society (ACS), 2021.
-
Abstract
- Cell-free DNA (cfDNA) analysis, specifically circulating tumor DNA (ctDNA) analysis, provides enormous opportunities for noninvasive early assessment of cancers. To date, PCR-based methods have led this field. However, the limited sensitivity/specificity of PCR-based methods necessitates the search for new methods. Here, we describe a direct approach to detect KRAS G12D mutated genes in clinical ctDNA samples with the utmost LOD and sensitivity/specificity. In this study, MutS protein was immobilized on the tip of an atomic force microscope (AFM), and the protein sensed the mismatched sites of the duplex formed between the capture probe on the surface and mutated DNA. A noteworthy LOD (3 copies, 0.006% allele frequency) was achieved, along with superb sensitivity/specificity (100%/100%). These observations demonstrate that force-based AFM, in combination with the protein found in nature and properly designed capture probes/blockers, represents an exciting new avenue for ctDNA analysis.
- Subjects :
- Atomic force microscopy
Mechanical Engineering
Point mutation
Bioengineering
General Chemistry
Condensed Matter Physics
medicine.disease_cause
Sensitivity and Specificity
Molecular biology
Circulating Tumor DNA
chemistry.chemical_compound
chemistry
Cell-free fetal DNA
Duplex (building)
Neoplasms
Mutation
Biomarkers, Tumor
medicine
Humans
Point Mutation
General Materials Science
KRAS
Gene
Allele frequency
DNA
Subjects
Details
- ISSN :
- 15306992 and 15306984
- Volume :
- 21
- Database :
- OpenAIRE
- Journal :
- Nano Letters
- Accession number :
- edsair.doi.dedup.....50fc38d0a6b0de71de0b85e7a8567ffb