Ultra-deep massively parallel sequencing with unique molecular identifier tagging achieves comparable performance to droplet digital PCR for detection and quantification of circulating tumor DNA from lung cancer patients.

The identification and quantification of actionable mutations are of critical importance for effective genotype-directed therapies, prognosis and drug response monitoring in patients with non-small-cell lung cancer (NSCLC). Although tumor tissue biopsy remains the gold standard for diagnosis of NSCL...

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Main Authors: Le Son Tran, Hong-Anh Thi Pham, Vu-Uyen Tran, Thanh-Truong Tran, Anh-Thu Huynh Dang, Dinh-Thong Le, Son-Lam Nguyen, Ngoc-Vu Nguyen, Trieu-Vu Nguyen, Binh Thanh Vo, Hong-Thuy Thi Dao, Nguyen Huu Nguyen, Tam Huu Tran, Chu Van Nguyen, Phuong Cam Pham, Anh Tuan Dang-Mai, Thien Kim Dinh-Nguyen, Van Hieu Phan, Thanh-Thuy Thi Do, Kiet Truong Dinh, Han Ngoc Do, Minh-Duy Phan, Hoa Giang, Hoai-Nghia Nguyen
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2019-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0226193&type=printable
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author Le Son Tran
Hong-Anh Thi Pham
Vu-Uyen Tran
Thanh-Truong Tran
Anh-Thu Huynh Dang
Dinh-Thong Le
Son-Lam Nguyen
Ngoc-Vu Nguyen
Trieu-Vu Nguyen
Binh Thanh Vo
Hong-Thuy Thi Dao
Nguyen Huu Nguyen
Tam Huu Tran
Chu Van Nguyen
Phuong Cam Pham
Anh Tuan Dang-Mai
Thien Kim Dinh-Nguyen
Van Hieu Phan
Thanh-Thuy Thi Do
Kiet Truong Dinh
Han Ngoc Do
Minh-Duy Phan
Hoa Giang
Hoai-Nghia Nguyen
author_facet Le Son Tran
Hong-Anh Thi Pham
Vu-Uyen Tran
Thanh-Truong Tran
Anh-Thu Huynh Dang
Dinh-Thong Le
Son-Lam Nguyen
Ngoc-Vu Nguyen
Trieu-Vu Nguyen
Binh Thanh Vo
Hong-Thuy Thi Dao
Nguyen Huu Nguyen
Tam Huu Tran
Chu Van Nguyen
Phuong Cam Pham
Anh Tuan Dang-Mai
Thien Kim Dinh-Nguyen
Van Hieu Phan
Thanh-Thuy Thi Do
Kiet Truong Dinh
Han Ngoc Do
Minh-Duy Phan
Hoa Giang
Hoai-Nghia Nguyen
author_sort Le Son Tran
collection DOAJ
description The identification and quantification of actionable mutations are of critical importance for effective genotype-directed therapies, prognosis and drug response monitoring in patients with non-small-cell lung cancer (NSCLC). Although tumor tissue biopsy remains the gold standard for diagnosis of NSCLC, the analysis of circulating tumor DNA (ctDNA) in plasma, known as liquid biopsy, has recently emerged as an alternative and noninvasive approach for exploring tumor genetic constitution. In this study, we developed a protocol for liquid biopsy using ultra-deep massively parallel sequencing (MPS) with unique molecular identifier tagging and evaluated its performance for the identification and quantification of tumor-derived mutations from plasma of patients with advanced NSCLC. Paired plasma and tumor tissue samples were used to evaluate mutation profiles detected by ultra-deep MPS, which showed 87.5% concordance. Cross-platform comparison with droplet digital PCR demonstrated comparable detection performance (91.4% concordance, Cohen's kappa coefficient of 0.85 with 95% CI = 0.72-0.97) and great reliability in quantification of mutation allele frequency (Intraclass correlation coefficient of 0.96 with 95% CI = 0.90-0.98). Our results highlight the potential application of liquid biopsy using ultra-deep MPS as a routine assay in clinical practice for both detection and quantification of actionable mutation landscape in NSCLC patients.
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spelling doaj-art-d2c70cc813ba446987362f652e3f3ffe2025-08-20T02:55:14ZengPublic Library of Science (PLoS)PLoS ONE1932-62032019-01-011412e022619310.1371/journal.pone.0226193Ultra-deep massively parallel sequencing with unique molecular identifier tagging achieves comparable performance to droplet digital PCR for detection and quantification of circulating tumor DNA from lung cancer patients.Le Son TranHong-Anh Thi PhamVu-Uyen TranThanh-Truong TranAnh-Thu Huynh DangDinh-Thong LeSon-Lam NguyenNgoc-Vu NguyenTrieu-Vu NguyenBinh Thanh VoHong-Thuy Thi DaoNguyen Huu NguyenTam Huu TranChu Van NguyenPhuong Cam PhamAnh Tuan Dang-MaiThien Kim Dinh-NguyenVan Hieu PhanThanh-Thuy Thi DoKiet Truong DinhHan Ngoc DoMinh-Duy PhanHoa GiangHoai-Nghia NguyenThe identification and quantification of actionable mutations are of critical importance for effective genotype-directed therapies, prognosis and drug response monitoring in patients with non-small-cell lung cancer (NSCLC). Although tumor tissue biopsy remains the gold standard for diagnosis of NSCLC, the analysis of circulating tumor DNA (ctDNA) in plasma, known as liquid biopsy, has recently emerged as an alternative and noninvasive approach for exploring tumor genetic constitution. In this study, we developed a protocol for liquid biopsy using ultra-deep massively parallel sequencing (MPS) with unique molecular identifier tagging and evaluated its performance for the identification and quantification of tumor-derived mutations from plasma of patients with advanced NSCLC. Paired plasma and tumor tissue samples were used to evaluate mutation profiles detected by ultra-deep MPS, which showed 87.5% concordance. Cross-platform comparison with droplet digital PCR demonstrated comparable detection performance (91.4% concordance, Cohen's kappa coefficient of 0.85 with 95% CI = 0.72-0.97) and great reliability in quantification of mutation allele frequency (Intraclass correlation coefficient of 0.96 with 95% CI = 0.90-0.98). Our results highlight the potential application of liquid biopsy using ultra-deep MPS as a routine assay in clinical practice for both detection and quantification of actionable mutation landscape in NSCLC patients.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0226193&type=printable
spellingShingle Le Son Tran
Hong-Anh Thi Pham
Vu-Uyen Tran
Thanh-Truong Tran
Anh-Thu Huynh Dang
Dinh-Thong Le
Son-Lam Nguyen
Ngoc-Vu Nguyen
Trieu-Vu Nguyen
Binh Thanh Vo
Hong-Thuy Thi Dao
Nguyen Huu Nguyen
Tam Huu Tran
Chu Van Nguyen
Phuong Cam Pham
Anh Tuan Dang-Mai
Thien Kim Dinh-Nguyen
Van Hieu Phan
Thanh-Thuy Thi Do
Kiet Truong Dinh
Han Ngoc Do
Minh-Duy Phan
Hoa Giang
Hoai-Nghia Nguyen
Ultra-deep massively parallel sequencing with unique molecular identifier tagging achieves comparable performance to droplet digital PCR for detection and quantification of circulating tumor DNA from lung cancer patients.
PLoS ONE
title Ultra-deep massively parallel sequencing with unique molecular identifier tagging achieves comparable performance to droplet digital PCR for detection and quantification of circulating tumor DNA from lung cancer patients.
title_full Ultra-deep massively parallel sequencing with unique molecular identifier tagging achieves comparable performance to droplet digital PCR for detection and quantification of circulating tumor DNA from lung cancer patients.
title_fullStr Ultra-deep massively parallel sequencing with unique molecular identifier tagging achieves comparable performance to droplet digital PCR for detection and quantification of circulating tumor DNA from lung cancer patients.
title_full_unstemmed Ultra-deep massively parallel sequencing with unique molecular identifier tagging achieves comparable performance to droplet digital PCR for detection and quantification of circulating tumor DNA from lung cancer patients.
title_short Ultra-deep massively parallel sequencing with unique molecular identifier tagging achieves comparable performance to droplet digital PCR for detection and quantification of circulating tumor DNA from lung cancer patients.
title_sort ultra deep massively parallel sequencing with unique molecular identifier tagging achieves comparable performance to droplet digital pcr for detection and quantification of circulating tumor dna from lung cancer patients
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0226193&type=printable
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