Benchmarking of Mutation Diagnostics in Clinical Lung Cancer Specimens

被引:96
作者
Querings, Silvia [1 ,2 ,3 ]
Altmueller, Janine [4 ]
Ansen, Sascha [5 ,6 ]
Zander, Thomas [5 ,6 ]
Seidel, Danila [1 ,2 ,3 ]
Gabler, Franziska [1 ,2 ,3 ]
Peifer, Martin [1 ,2 ]
Markert, Eva [7 ]
Stemshorn, Kathryn [4 ]
Timmermann, Bernd [8 ]
Saal, Beate [1 ,2 ]
Klose, Stefan [10 ]
Ernestus, Karen [7 ]
Scheffler, Matthias [5 ,6 ]
Engel-Riedel, Walburga [9 ]
Stoelben, Erich [11 ]
Brambilla, Elisabeth [12 ]
Wolf, Juergen [3 ,5 ,6 ]
Nuernberg, Peter [4 ]
Thomas, Roman K. [1 ,2 ,3 ,5 ,6 ]
机构
[1] Max Planck Gesell, Klaus Joachim Zulch Labs, Max Planck Inst Neurol Res, Cologne, Germany
[2] Univ Cologne, Fac Med, Cologne, Germany
[3] Univ Cologne, Ctr Integrated Oncol Koln Bonn, Lab Translat Canc Genom, Cologne, Germany
[4] Univ Cologne, Cologne Ctr Genom, Cologne, Germany
[5] Univ Hosp Cologne, Dept Internal Med 1, Cologne, Germany
[6] Univ Hosp Cologne, Ctr Integrated Oncol Koln Bonn, Cologne, Germany
[7] Hosp Merheim, Inst Pathol, Cologne, Germany
[8] Max Planck Inst Mol Genet, Berlin, Germany
[9] Lung Clin Merheim, Dept Med Oncol, Cologne, Germany
[10] Pneumol Gemeinschaftpraxis Dr HP Wagner Dr St Kos, Chemnitz, Germany
[11] Lung Clin Merheim, Dept Thorac Surg, Cologne, Germany
[12] Univ Grenoble 1, CHU Grenoble Hop Michallon, INSERM, Dept Pathol,Inst Albert Bonniot,U823, Grenoble, France
来源
PLOS ONE | 2011年 / 6卷 / 05期
关键词
GROWTH-FACTOR RECEPTOR; EGFR MUTATIONS; GENE-MUTATIONS; KRAS MUTATION; GEFITINIB; POLYMERASE; SYSTEM; SAMPLES; TUMORS;
D O I
10.1371/journal.pone.0019601
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Treatment of EGFR-mutant non-small cell lung cancer patients with the tyrosine kinase inhibitors erlotinib or gefitinib results in high response rates and prolonged progression-free survival. Despite the development of sensitive mutation detection approaches, a thorough validation of these in a clinical setting has so far been lacking. We performed, in a clinical setting, a systematic validation of dideoxy 'Sanger' sequencing and pyrosequencing against massively parallel sequencing as one of the most sensitive mutation detection technologies available. Mutational annotation of clinical lung tumor samples revealed that of all patients with a confirmed response to EGFR inhibition, only massively parallel sequencing detected all relevant mutations. By contrast, dideoxy sequencing missed four responders and pyrosequencing missed two responders, indicating a dramatic lack of sensitivity of dideoxy sequencing, which is widely applied for this purpose. Furthermore, precise quantification of mutant alleles revealed a low correlation (r(2) = 0.27) of histopathological estimates of tumor content and frequency of mutant alleles, thereby questioning the use of histopathology for stratification of specimens for individual analytical procedures. Our results suggest that enhanced analytical sensitivity is critically required to correctly identify patients responding to EGFR inhibition. More broadly, our results emphasize the need for thorough evaluation of all mutation detection approaches against massively parallel sequencing as a prerequisite for any clinical implementation.
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页数:8
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