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Related Experiment Video

Updated: Oct 14, 2025

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Published on: March 29, 2017

A Sensitive PCR-Based Method for Somatic Mutations Enrichment and Screening.

Yaming Xiong¹, Hailing Tang¹

¹Department of Thoracic Surgery, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510120, People's Republic of China.

Cancer Management and Research

|November 5, 2021

Summary

A novel nuclease-based assay enriches mutant DNA from plasma, achieving 0.001% sensitivity for EGFR and KRAS mutations in lung cancer screening. This cost-effective method improves noninvasive cancer monitoring.

Keywords:

Surveyor enrichment liquid biopsy lung cancer

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Last Updated: Oct 14, 2025

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Published on: March 29, 2017

Author Spotlight: Advancing the Detection of Low-Frequency Mutations in Cancer Tissues

Author Spotlight: Advancing the Detection of Low-Frequency Mutations in Cancer Tissues

Published on: August 23, 2024

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Published on: September 25, 2018

Area of Science:

Oncology
Molecular Biology
Genetics

Background:

Epidermal Growth Factor Receptor (EGFR) and KRAS are the most frequently mutated genes in lung cancers, present in approximately 60% of cases.
Mutation gene assays are promising noninvasive biomarkers for monitoring cancer dynamics.
Detecting low levels of tumor-derived DNA fragments in plasma presents a significant challenge.

Purpose of the Study:

To develop a sensitive and efficient method for enriching mutant DNA from plasma for cancer screening.
To validate a nuclease-based assay for detecting EGFR and KRAS mutations.

Main Methods:

A nuclease-based enrichment assay utilizing Surveyor endonuclease to cleave mismatched DNA molecules was developed.
The assay was used to screen lung cancer specimens for mutations in EGFR exons 18 and 21, and KRAS exon 2 (codons 12 and 13).
A single set of primers and three TaqMan probes were employed for screening all target mutant genes.

Main Results:

The method effectively removes wild-type sequences and enriches mutation DNA.
A minimum detectable mutant allele frequency (MAF) of 0.001% was achieved, demonstrating high sensitivity.
The assay demonstrated increased sensitivity and efficiency for mutation DNA detection in cancer screening.

Conclusions:

A simple, sensitive, and cost-effective methodology for mutation gene screening was developed.
The nuclease-based enrichment assay is suitable for detecting complex DNA variations like EGFR and KRAS mutations.
This method enhances the potential for noninvasive cancer monitoring through blood-based biomarkers.