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首页> 外文期刊>Pathology Research and Practice >Frequency of KRAS, BRAF, and PIK3CA mutations in advanced colorectal cancers: Comparison of peptide nucleic acid-mediated PCR clamping and direct sequencing in formalin-fixed, paraffin-embedded tissue.
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Frequency of KRAS, BRAF, and PIK3CA mutations in advanced colorectal cancers: Comparison of peptide nucleic acid-mediated PCR clamping and direct sequencing in formalin-fixed, paraffin-embedded tissue.

机译：晚期结直肠癌中KRAS，BRAF和PIK3CA突变的频率：在福尔马林固定，石蜡包埋的组织中比较肽核酸介导的PCR钳制和直接测序。

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摘要

KRAS, BRAF, and PIK3CA mutation testing before administration of anti-epidermal growth factor receptor therapy of metastatic colorectal cancer (CRC) has become important. However, considerable uncertainty exists regarding which detection method can be applied in a reproducible, sensitive, and simple manner in the routine diagnostic setting. We compared the detection rates of KRAS, BRAF, and PIK3CA mutations in 92 routine formalin-fixed, paraffin-embedded CRC specimens by 2 discrete methods: direct sequencing and peptide nucleic acid (PNA)-mediated PCR. The detection rates for KRAS, BRAF, and PIK3CA mutations by direct sequencing were 20.7%, 3.3%, and 1.1%, respectively. PNA-mediated PCR clamping significantly increased the percentages of KRAS, BRAF, and PIK3CA mutations by up to 7.6%, 1.2%, and 5.4%, respectively, compared to the detection rate of regular PCR followed by direct sequencing (p=0.039, p=0.250, and p=0.031, respectively). The tumor volume of discordant cases was not significantly different from concordant cases (56.2+/-28.7% vs. 67.6+/-17.9%, p=0.41), which implies that there is a minor population of mutant alleles in the heterogeneous tumor population. The PNA-mediated PCR clamping method is highly sensitive and is efficiently applicable to the detection of KRAS, BRAF, and PIK3CA mutations in a clinical setting.
机译：施用抗表皮生长因子受体治疗转移性结直肠癌（CRC）之前的KRAS，BRAF和PIK3CA突变测试变得非常重要。然而，关于在常规诊断设置中可以以可再现，灵敏和简单的方式应用哪种检测方法，存在很大的不确定性。我们比较了92种常规福尔马林固定，石蜡包埋的CRC标本中KRAS，BRAF和PIK3CA突变的检测率，该方法通过2种离散方法：直接测序和肽核酸（PNA）介导的PCR。通过直接测序对KRAS，BRAF和PIK3CA突变的检出率分别为20.7％，3.3％和1.1％。与常规PCR和直接测序的检测率相比，PNA介导的PCR钳制显着提高了KRAS，BRAF和PIK3CA突变的百分比，分别高达7.6％，1.2％和5.4％（p = 0.039，p = 0.250和p = 0.031）。不一致病例的肿瘤体积与一致病例无显着差异（56.2 +/- 28.7％vs. 67.6 +/- 17.9％，p = 0.41），这表明异质性肿瘤群体中存在少量突变等位基因。 PNA介导的PCR钳制方法高度灵敏，可有效应用于临床环境中KRAS，BRAF和PIK3CA突变的检测。

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