Related Experiment Video
Updated: May 10, 2026
10:57
Visualizing Genetic Variants, Short Targets, and Point Mutations in the Morphological Tissue Context with an RNA In Situ Hybridization Assay
Published on: August 14, 2018
DISSECT Method Using PNA-LNA Clamp Improves Detection of EGFR T790m Mutation
Minakshi Guha1, Elena Castellanos-Rizaldos, G Mike Makrigiorgos
1Division of DNA Repair and Genome Stability, Department of Radiation Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts, United States of America.
Plos One
|June 28, 2013
Summary
A new method combines DISSECT and PNA-LNA PCR to detect low-level EGFR T790M mutations in non-small cell lung cancer (NSCLC). This advance aids in identifying resistance mutations crucial for treatment monitoring.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Non-small cell lung cancer (NSCLC) patients often develop resistance to EGFR inhibitors via secondary mutations, notably T790M.
- These resistance mutations can arise from small subclonal populations within the primary tumor.
- Sensitive detection methods are crucial for identifying low-level DNA variations in tumors and plasma.
Purpose of the Study:
- To develop and validate a highly sensitive method for detecting the T790M EGFR mutation.
- To combine Differential Strand Separation at Critical Temperature (DISSECT) with PNA-LNA PCR for enhanced mutation detection.
- To enable early detection of resistance mechanisms in NSCLC.
Main Methods:
- Utilized Differential Strand Separation at Critical Temperature (DISSECT) for pre-enrichment of low-abundance T790M EGFR mutations.
- Employed peptide nucleic acid-locked nucleic acid (PNA-LNA) polymerase chain reaction (PCR) for sensitive mutation detection.
- Combined DISSECT with PNA-LNA PCR to achieve a detection limit of 1 mutant allele in 10,000 wild-type alleles.
Main Results:
- The combined DISSECT-PNA-LNA PCR methodology demonstrated superior sensitivity in detecting T790M EGFR mutations.
- Achieved detection of 1 mutant allele against a background of 10,000 wild-type alleles.
- Established a robust method for identifying low-frequency resistance mutations.
Conclusions:
- The DISSECT-PNA-LNA PCR combination offers a highly sensitive approach for detecting T790M EGFR mutations.
- This methodology is adaptable for the sensitive detection of other emerging cancer resistance mutations.
- Enables improved monitoring of treatment resistance in NSCLC patients.
