Multiplex Digital Methylation-Specific PCR for Noninvasive Screening of Lung Cancer.

Autor:	Zhao Y; Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD, 21287, USA., O'Keefe CM; Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD, 21287, USA., Hsieh K; Department of Mechanical Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA., Cope L; Department of Oncology, Johns Hopkins University, Baltimore, MD, 21287, USA., Joyce SC; The UPMC Hillman Cancer Center, University of Pittsburgh, Pittsburgh, PA, 15232, USA.; Division of Hematology and Oncology, Department of Medicine, University of Pittsburgh Medical Center, Pittsburgh, PA, United States., Pisanic TR; Department of Oncology, Johns Hopkins University, Baltimore, MD, 21287, USA.; Johns Hopkins Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA., Herman JG; The UPMC Hillman Cancer Center, University of Pittsburgh, Pittsburgh, PA, 15232, USA.; Division of Hematology and Oncology, Department of Medicine, University of Pittsburgh Medical Center, Pittsburgh, PA, United States., Wang TH; Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD, 21287, USA.; Department of Mechanical Engineering, Johns Hopkins University, Baltimore, MD, 21218, USA.; Johns Hopkins Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, 21218, USA.
Jazyk:	angličtina
Zdroj:	Advanced science (Weinheim, Baden-Wurttemberg, Germany) [Adv Sci (Weinh)] 2023 Jun; Vol. 10 (16), pp. e2206518. Date of Electronic Publication: 2023 Apr 11.
DOI:	10.1002/advs.202206518
Abstrakt:	There remains tremendous interest in developing liquid biopsy assays for detection of cancer-specific alterations, such as mutations and DNA methylation, in cell-free DNA (cfDNA) obtained through noninvasive blood draws. However, liquid biopsy analysis is often challenging due to exceedingly low fractions of circulating tumor DNA (ctDNA), necessitating the use of extended tumor biomarker panels. While multiplexed PCR strategies provide advantages such as higher throughput, their implementation is often hindered by challenges such as primer-dimers and PCR competition. Alternatively, digital PCR (dPCR) approaches generally offer superior performance, but with constrained multiplexing capability. This paper describes development and validation of the first multiplex digital methylation-specific PCR (mdMSP) platform for simultaneous analysis of four methylation biomarkers for liquid-biopsy-based detection of non-small cell lung cancer (NSCLC). mdMSP employs a microfluidic device containing four independent, but identical modules, housing a total of 40 160 nanowells. Analytical validation of the mdMSP platform demonstrates multiplex detection at analytical specificities as low as 0.0005%. The clinical utility of mdMSP is also demonstrated in a cohort of 72 clinical samples of low-volume liquid biopsy specimens from patients with computed tomography (CT)-scan indeterminant pulmonary nodules, exhibiting superior clinical performance when compared to traditional MSP assays for noninvasive detection of early-stage NSCLC. (© 2023 The Authors. Advanced Science published by Wiley-VCH GmbH.)
Databáze:	MEDLINE
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