Tissue and Plasma-Based Highly Sensitive Blocker Displacement Amplicon Nanopore Sequencing for EGFR Mutations in Lung Cancer
2
Issued Date
2024-04-01
Resource Type
ISSN
15982998
eISSN
20059256
Scopus ID
2-s2.0-85190338606
Pubmed ID
37986562
Journal Title
Cancer Research and Treatment
Volume
56
Issue
2
Start Page
455
End Page
463
Rights Holder(s)
SCOPUS
Bibliographic Citation
Cancer Research and Treatment Vol.56 No.2 (2024) , 455-463
Suggested Citation
Akkhasutthikun P., Kaewsapsak P., Nimsamer P., Klomkliew P., Visedthorn S., Chanchaem P., Teerapakpinyo C., Payungporn S., Luangdilok S. Tissue and Plasma-Based Highly Sensitive Blocker Displacement Amplicon Nanopore Sequencing for EGFR Mutations in Lung Cancer. Cancer Research and Treatment Vol.56 No.2 (2024) , 455-463. 463. doi:10.4143/crt.2023.1108 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/98065
Title
Tissue and Plasma-Based Highly Sensitive Blocker Displacement Amplicon Nanopore Sequencing for EGFR Mutations in Lung Cancer
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Corresponding Author(s)
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Abstract
Purpose The epidermal growth factor receptor (EGFR) mutation is a widely prevalent oncogene driver in non–small cell lung cancer (NSCLC) in East Asia. The detection of EGFR mutations is a standard biomarker test performed routinely in patients with NSCLC for the selection of targeted therapy. Here, our objective was to develop a portable new technique for detecting EGFR (19Del, T790M, and L858R) mutations based on Nanopore sequencing. Materials and Methods The assay employed a blocker displacement amplification (BDA)–based polymerase chain reaction (PCR) technique combined with Nanopore sequencing to detect EGFR mutations. Mutant and wild-type EGFR clones were generated from DNA from H1650 (19Del heterozygous) and H1975 (T790M and L858R heterozygous) lung cancer cell lines. Then, they were mixed to assess the performance of this technique for detecting low variant allele frequencies (VAFs). Subsequently, formalin-fixed, paraffin-embedded (FFPE) tissue and cell-free DNA (cfDNA) from patients with NSCLC were used for clinical validation. Results The assay can detect low VAF at 0.5% mutant mixed in wild-type EGFR. Using FFPE DNA, the concordance rates of EGFR 19Del, T790M, and L858R mutations between our method and Cobas real-time PCR were 98.46%, 100%, and 100%, respectively. For cfDNA, the concordance rates of EGFR 19Del, T790M, and L858R mutations between our method and droplet digital PCR were 94.74%, 100%, and 100%, respectively. Conclusion The BDA amplicon Nanopore sequencing is a highly accurate and sensitive method for the detection of EGFR mutations in clinical specimens.