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Updated: Jul 15, 2026

Integration of Wet and Dry Bench Processes Optimizes Targeted Next-generation Sequencing of Low-quality and Low-quantity Tumor Biopsies

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Published on: April 11, 2016

Development and validation of an ultrasensitive qPCR method to identify and quantify EGFR T790M in cell-free DNA

Shenglei Yuan¹, Nan Jia¹, Guofu Lu¹

¹Bioanalytical Services Department, WuXi AppTec (Shanghai) Co. Ltd, Shanghai, China.

|January 15, 2025

Abstract

Background:

Circulating tumor DNA (ctDNA) is a promising biomarker for cancer prognosis and drug development. A major challenge in the ctDNA determination method is discriminating ctDNA from highly similar but significantly more abundant wild-type DNA sensitively and accurately.

Method:

An ultrasensitive qPCR method termed Triple Enrichment Amplification of Mutation PCR (TEAM-PCR) was developed to detect EGFR T790M mutation.

Results:

EGFR T790M was quantified over the assay range of 25-10⁶ copies/reaction in the presence of 10⁶ wild-type copies. This method was fully validated following the essential bioanalysis guidance, with the limit of detection (LOD) being five copies/reaction.

Conclusion:

This study established and validated a qPCR-based strategy to detect EGFR T790M mutation with ultra-high sensitivity and reliability.

Keywords:

EGFR T790M LOD NSCLC TEAM-PCR bioanalysis ctDNA method validation point mutation qPCR sensitivity

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Establishment and Characterization of Three Afatinib-resistant Lung Adenocarcinoma PC-9 Cell Lines Developed with Increasing Doses of Afatinib

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The real-time quantification of the number of amplified products is...

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