Highly Sensitive 3D‐Nanoplasmonic‐Based Epidermal Growth Factor Receptor Mutation Multiplex Assay Chip for Liquid Biopsy

Author:

Lee Ji Young1,Jeong Byeong‐Ho2,Jung Ho Sang1,Kang Taejoon34,Park Yeonkyung3,Rho Jin Kyung5,Park Sung‐Gyu1ORCID,Lee Min‐Young1ORCID

Affiliation:

1. Advanced Bio and Healthcare Materials Research Division Korea Institute of Materials Science (KIMS) 797, Changwon‐daero, Seongsan‐gu Changwon‐si Gyeongsangnam‐do 51508 Republic of Korea

2. Division of Pulmonary and Critical Care Medicine Department of Medicine Samsung Medical Center SungkyunKwan University School of Medicine 81, Irwon‐ro, Gangnam‐gu Seoul 06351 Republic of Korea

3. Bionanotechnology Research Center Korea Research Institute of Bioscience and Biotechnology (KRIBB) 125 Gwahak‐ro, Yuseong‐gu Daejeon 34141 Republic of Korea

4. School of Pharmacy Sungkyunkwan University 2066 Seobu‐ro, Jangan‐gu Suwon, Gyeonggi‐do 16419 Republic of Korea

5. Department of Convergence Medicine Asan Medical Center College of Medicine University of Ulsan Olympic‐ro 43‐gil, Songpa‐gu Seoul 05505 Republic of Korea

Abstract

Economical mutation detection method with high analytical and clinical sensitivity is necessary for early cancer diagnosis and screening. In this study, a novel 3D‐nanoplasmonic‐based multiplex mutation assay chip is developed to detect epidermal growth factor receptor (EGFR) mutations. This assay kit comprises a 3D‐nanoplasmonic substrate immobilized with capture probes and primer–probe sets for recombinase polymerase amplification, wild‐type inhibition, and fluorescence detection, enabling multiplex detection of EGFR exon 19 deletions, exon 20 insertions, and exon 21 L858R point mutations. The strategy facilitates the detection of all deletions and insertions within the target region with extremely high analytical sensitivity, detecting as low as 1 × 10−9% mutation frequency, implying three copies/reactions and 100 zM. The synergistic effects of plasmon‐enhanced fluorescence from the 3D‐nanoplasmonic substrate and wild‐type inhibitor contribute to this high analytical sensitivity. Moreover, the developed chip exhibits 100% accuracy in the clinical testing of plasma samples from normal individuals and patients with benign lung tumor and malignant lung tumor. With high sensitivity and multiplexing capabilities, this assay operates at a low reaction temperature (around 37 °C) and requires a short processing time, ≈70 min post‐cell‐free DNA extraction. These features make the chip a valuable tool for easy and widespread cancer screening.

Publisher

Wiley

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