Multiplex Digital Methylation-Specific PCR for Noninvasive Screening of Lung Cancer.
Multiplex Digital Methylation-Specific PCR for Noninvasive Screening of Lung Cancer.
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DOI：
10.1002/advs.202206518
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发表时间：
2023-06
期刊：
影响因子：
15.1
通讯作者：
Wang, Tza-Huei
中科院分区：
文献类型：
--
作者：
Zhao, Yang;O'Keefe, Christine M.;Hsieh, Kuangwen;Cope, Leslie;Joyce, Sonali C.;Pisanic, Thomas R.;Herman, James G.;Wang, Tza-Huei
在开发用于检测癌症特异性改变的液体活检测定方面仍然存在巨大的兴趣，例如通过非侵入性抽血获得的无细胞DNA（cfDNA）中的突变和DNA甲基化。然而，由于循环肿瘤DNA（ctDNA）的分数极低，液体活检分析通常具有挑战性，需要使用扩展的肿瘤生物标志物组。虽然多重PCR策略提供了诸如更高通量等优点，但它们的实施通常受到诸如引物二聚体和PCR竞争等挑战的阻碍。或者，数字PCR（dPCR）方法通常提供上级性能，但具有受限的多路复用能力。本文描述了第一个多重数字甲基化特异性PCR（mdMSP）平台的开发和验证，该平台用于同时分析四种甲基化生物标志物，用于基于液体活检的非小细胞肺癌（NSCLC）检测。mdMSP采用包含四个独立但相同的模块的微流体装置，容纳总共40 160个微细胞。mdMSP平台的分析验证表明，多重检测的分析特异性低至0.0005%。mdMSP的临床效用也在来自计算机断层扫描（CT）扫描不确定性肺结节患者的72份低体积液体活检标本的临床样本队列中得到证实，与传统MSP检测相比，其在早期NSCLC无创检测中表现出上级临床性能。本文描述了第一个mdMSP平台的开发和分析/临床验证，该平台用于同时分析四种DNA甲基化生物标志物，用于基于液体活检的早期NSCLC检测。所展示的分析灵敏度/特异性和临床实用性表明mdMSP平台有潜力成为改善肺癌CT筛查的补充方法。
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. This paper describes the development and analytical/clinical validation of the first mdMSP platform for the simultaneous analysis of four DNA methylation biomarkers for liquid‐biopsy‐based detection of early‐stage NSCLC. The demonstrated analytical sensitivity/specificity and clinical utility indicate the potential of mdMSP platform to be a complementary method for improved CT screening of lung cancer.
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影响因子：
5.7
作者：
Guo S;Yan F;Xu J;Bao Y;Zhu J;Wang X;Wu J;Li Y;Pu W;Liu Y;Jiang Z;Ma Y;Chen X;Xiong M;Jin L;Wang J
通讯作者：
Wang J
影响因子：
7
作者：
Amelio I;Bertolo R;Bove P;Buonomo OC;Candi E;Chiocchi M;Cipriani C;Di Daniele N;Ganini C;Juhl H;Mauriello A;Marani C;Marshall J;Montanaro M;Palmieri G;Piacentini M;Sica G;Tesauro M;Rovella V;Tisone G;Shi Y;Wang Y;Melino G
通讯作者：
Melino G
DOI：
10.1073/pnas.93.18.9821
发表时间：
1996-09-03
影响因子：
11.1
作者：
Herman, JG;Graff, JR;Baylin, SB
通讯作者：
Baylin, SB
影响因子：
158.5
作者：
Brock, Malcolm V.;Hooker, Craig M.;Herman, James G.
通讯作者：
Herman, James G.
影响因子：
37.3
作者：
Anglim, Paul P.;Alonzo, Todd A.;Laird-Offringa, Ite A.
通讯作者：
Laird-Offringa, Ite A.