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Published on: August 14, 2018
A mutation-sensitive switch assay to detect five clinically significant epidermal growth factor receptor mutations
Bin Liu1, Lin Zhou1, Qian Wang1
1Department of Molecular Diagnostics and Biopharmaceutics, College of Pharmaceutical Science, Soochow University , Suzhou, China .
Abstract:
Epidermal growth factor receptor (EGFR) mutations can affect the therapeutic efficacy of drugs used to treat nonsmall-cell lung cancer (NSCLC). We aimed to develop methods to detect five common EGFR somatic mutations in tumor tissues from NSCLC patients by using a nanoscale mutation-sensitive switch consisting of a high-fidelity polymerase and phosphorothioate-modified allele-specific primers. The five clinically significant EGFR mutations examined here are S768I, T790M, L858R, and 15- and 18-bp deletion mutations in exon 19. Our assays showed sensitivities of 100 copies and specificities of more than three log scales for matched templates relative to mismatched templates by routine polymerase chain reaction (PCR), real-time PCR, and multiplex PCR. This assay would be superior to DNA sequencing in situations where mutant DNA is not abundant.
Insights
This study presents a novel nanoscale assay for detecting common epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer (NSCLC). The method offers high sensitivity and specificity, outperforming DNA sequencing for low-abundance mutant DNA.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Epidermal growth factor receptor (EGFR) mutations are critical in non-small cell lung cancer (NSCLC) therapeutics.
- Accurate detection of these mutations impacts treatment efficacy.
Purpose of the Study:
- To develop a sensitive and specific method for detecting five common EGFR somatic mutations in NSCLC tumor tissues.
- To evaluate the performance of a nanoscale mutation-sensitive switch assay.
Main Methods:
- Utilized a nanoscale mutation-sensitive switch with high-fidelity polymerase and allele-specific primers.
- Tested the assay using routine PCR, real-time PCR, and multiplex PCR.
- Included five clinically significant EGFR mutations: S768I, T790M, L858R, and exon 19 deletions (15- and 18-bp).
Main Results:
- Achieved high sensitivity (100 copies) and specificity (over three log scales) for matched vs. mismatched templates.
- Demonstrated superior performance compared to DNA sequencing when mutant DNA is scarce.
Conclusions:
- The developed nanoscale assay effectively detects common EGFR mutations in NSCLC.
- This method provides a valuable alternative to DNA sequencing, especially for samples with low mutant DNA levels.
