Development of a highly sensitive liquid biopsy platform to detect clinically-relevant cancer mutations at low allele fractions in cell-free DNA.
Development of a highly sensitive liquid biopsy platform to detect clinically-relevant cancer mutations at low allele fractions in cell-free DNA.
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DOI：
10.1371/journal.pone.0194630
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Rosenfeld N
中科院分区：
文献类型：
--
作者：
Gale D;Lawson ARJ;Howarth K;Madi M;Durham B;Smalley S;Calaway J;Blais S;Jones G;Clark J;Dimitrov P;Pugh M;Woodhouse S;Epstein M;Fernandez-Gonzalez A;Whale AS;Huggett JF;Foy CA;Jones GM;Raveh-Amit H;Schmitt K;Devonshire A;Green E;Forshew T;Plagnol V;Rosenfeld N
循环肿瘤DNA（ctDNA）的检测和监测正迅速成为癌症患者护理中的诊断、预后和预测工具。越来越多的基因靶点已被确定为诊断性或可操作的，需要开发可靠的技术，提供并行的多个基因的分析。我们开发了InVision™液体活检平台，该平台利用增强型TAm-Seq™（eTAm-Seq™）技术，这是一种基于扩增子的下一代测序方法，用于在一组35个癌症相关基因中鉴定ctDNA中低频的临床相关体细胞改变。我们在两个实验室中对eTAm-Seq技术进行了分析验证，以确定突变鉴定的重现性。我们使用既定的DNA标准品和新型全过程对照材料，评估了eTAm-Seq技术与数字PCR（dPCR）相比分析临床相关基因单核苷酸变体的定量性能。该测定法检测到低至0.02% AF的突变等位基因，每碱基特异性高达99.9997%。在两个实验室中，对具有最佳DNA量的样品进行分析，在0.25%-0.33%等位基因分数（AF）下检测到94%的突变，对于具有较低输入DNA量的样品检测到90%的突变。这些研究表明，eTAm-Seq技术是一种用于鉴定和定量循环肿瘤DNA中体细胞突变的稳健且可重复的技术，并支持其在精准医学的临床应用中的使用。
Detection and monitoring of circulating tumor DNA (ctDNA) is rapidly becoming a diagnostic, prognostic and predictive tool in cancer patient care. A growing number of gene targets have been identified as diagnostic or actionable, requiring the development of reliable technology that provides analysis of multiple genes in parallel. We have developed the InVision™ liquid biopsy platform which utilizes enhanced TAm-Seq™ (eTAm-Seq™) technology, an amplicon-based next generation sequencing method for the identification of clinically-relevant somatic alterations at low frequency in ctDNA across a panel of 35 cancer-related genes. We present analytical validation of the eTAm-Seq technology across two laboratories to determine the reproducibility of mutation identification. We assess the quantitative performance of eTAm-Seq technology for analysis of single nucleotide variants in clinically-relevant genes as compared to digital PCR (dPCR), using both established DNA standards and novel full-process control material. The assay detected mutant alleles down to 0.02% AF, with high per-base specificity of 99.9997%. Across two laboratories, analysis of samples with optimal amount of DNA detected 94% mutations at 0.25%-0.33% allele fraction (AF), with 90% of mutations detected for samples with lower amounts of input DNA. These studies demonstrate that eTAm-Seq technology is a robust and reproducible technology for the identification and quantification of somatic mutations in circulating tumor DNA, and support its use in clinical applications for precision medicine.
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影响因子：
12.3
作者：
Park G;Park JK;Shin SH;Jeon HJ;Kim NKD;Kim YJ;Shin HT;Lee E;Lee KH;Son DS;Park WY;Park D
通讯作者：
Park D
影响因子：
4.4
作者：
Aigrain L;Gu Y;Quail MA
通讯作者：
Quail MA
DOI：
10.1056/nejmoa1409405
发表时间：
2014-12-25
期刊：
The New England journal of medicine
影响因子：
--
作者：
Genovese G;Kähler AK;Handsaker RE;Lindberg J;Rose SA;Bakhoum SF;Chambert K;Mick E;Neale BM;Fromer M;Purcell SM;Svantesson O;Landén M;Höglund M;Lehmann S;Gabriel SB;Moran JL;Lander ES;Sullivan PF;Sklar P;Grönberg H;Hultman CM;McCarroll SA
通讯作者：
McCarroll SA
影响因子：
12.3
作者：
McLaren W;Gil L;Hunt SE;Riat HS;Ritchie GR;Thormann A;Flicek P;Cunningham F
通讯作者：
Cunningham F
影响因子：
4.3
作者：
Devonshire, Alison S.;Whale, Alexandra S.;Gutteridge, Alice;Jones, Gerwyn;Cowen, Simon;Foy, Carole A.;Huggett, Jim F.
通讯作者：
Huggett, Jim F.