Benchmarking of mutation diagnostics in clinical lung cancer specimens.
Benchmarking of mutation diagnostics in clinical lung cancer specimens.
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DOI：
10.1371/journal.pone.0019601
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发表时间：
2011-05-05
期刊：
影响因子：
3.7
通讯作者：
Thomas RK
中科院分区：
文献类型：
--
作者：
Querings S;Altmüller J;Ansén S;Zander T;Seidel D;Gabler F;Peifer M;Markert E;Stemshorn K;Timmermann B;Saal B;Klose S;Ernestus K;Scheffler M;Engel-Riedel W;Stoelben E;Brambilla E;Wolf J;Nürnberg P;Thomas RK
使用酪氨酸激酶抑制剂厄洛替尼或吉非替尼治疗egfr突变的非小细胞肺癌患者可获得高缓解率和延长无进展生存期。尽管开发了敏感的突变检测方法，但迄今为止还缺乏在临床环境中对这些方法进行彻底验证。在临床环境中，我们对双脱氧Sanger测序和焦磷酸测序进行了系统验证，以对抗大规模平行测序作为最敏感的突变检测技术之一。临床肺肿瘤样本的突变注释显示，在所有证实对EGFR抑制有反应的患者中，只有大规模平行测序检测到所有相关突变。相比之下，二脱氧测序遗漏了4个应答者，焦磷酸测序遗漏了2个应答者，这表明二脱氧测序的灵敏度明显不足，而二脱氧测序被广泛用于这一目的。此外，突变等位基因的精确定量显示，肿瘤含量与突变等位基因频率的组织病理学估计相关性较低（r2 = 0.27），从而质疑组织病理学在个体分析过程中对标本分层的使用。我们的研究结果表明，提高分析灵敏度对于正确识别对EGFR抑制有反应的患者至关重要。更广泛地说，我们的结果强调需要对所有针对大规模并行测序的突变检测方法进行彻底评估，作为任何临床实施的先决条件。
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 (r2 = 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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影响因子：
158.5
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