Comprehensive Validation of Snapback Primer-Based Melting Curve Analysis to Detect Nucleotide Variation in the Codon 12 and 13 of KRAS Gene.
Comprehensive Validation of Snapback Primer-Based Melting Curve Analysis to Detect Nucleotide Variation in the Codon 12 and 13 of KRAS Gene.
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全面验证基于 Snapback 引物的熔解曲线分析以检测 KRAS 基因密码子 12 和 13 中的核苷酸变异。
DOI：
10.1155/2018/8727941
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发表时间：
2018
影响因子：
--
通讯作者：
Wu Z
中科院分区：
文献类型：
--
作者：
Zhang Q;Du Y;Zhang X;Kang Z;Guan M;Wu Z
肿瘤标本中KRAS基因分型是抗EGFR治疗管理的决定性临床检验。然而，不同肿瘤类型间KRAS基因突变格局的复杂性，以及肿瘤细胞性和异质性引起的马赛克效应，使得KRAS基因分型方法的选择成为临床实践中一个具有挑战性的课题。我们使用癌症基因组图谱(TCGA)和可操作癌症靶点综合突变图谱(MSKCC-IMPACT)数据库，分别描绘了30多种癌症中7844例原发肿瘤和10,336例转移瘤的体细胞KRAS突变情况。建立了一种基于熔化曲线分析的Snapback引物方法，用于检测KRAS密码子12和13中最常见的体细胞突变。通过对100例结直肠癌(CRC)标本进行基因分型，并与Sanger测序和T-A克隆测序进行比较，验证了该方法的敏感性和准确性。胰腺癌(体细胞突变率为90.6%)、结直肠癌(42.5%)和肺腺癌(32.6%)是KRAS基因突变最多的三种原发肿瘤类型。转移性肿瘤中KRAS基因突变的发生率(90.99%)和多样性(66.31%)均高于原发性肿瘤。密码子12和13的突变是KRAS的主要遗传改变(TCGA占84.15%，MSK-IMPACT占86.13%)。此外，KRAS突变与转移性癌症患者的总体生存高度相关。与Sanger测序法(突变载量20%，假阴性率7%)相比，Snapback引物法对KRAS基因第12、13密码子突变(1%突变载量)具有更好的富集性和检测性。KRAS突变模式在不同的癌症类型中差异很大，与转移性癌症患者的生存有关。Snapback法是一种可靠、灵敏的检测KRAS主要突变等位基因的方法，可能有助于有效的癌症治疗决策。
KRAS genotyping in tumor samples is a decisive clinical test for the anti-EGFR therapy management. However, the complexity of KRAS mutation landscape across different cancer types and the mosaic effect caused by cancer cellularity and heterogeneity make the choice of KRAS genotyping method a challenging topic in the clinical practice. We depicted the landscape of somatic KRAS mutation in 7,844 primary tumors and 10,336 metastatic tumors across over 30 types of cancer using the Cancer Genome Atlas (TCGA) and Integrated Mutation Profiling of Actionable Cancer Targets (MSKCC-IMPACT) databases, respectively. A snapback primer assay based on melting curve analysis was developed to detect the most common somatic mutations in KRAS codons 12 and 13. The sensitivity and accuracy of the method was validated by genotyping 100 colorectal cancer (CRC) samples, in comparison with Sanger sequencing and T-A cloning sequencing. Pancreas adenocarcinoma (somatic mutation frequency 90.6%), colorectal adenocarcinoma (42.5%), and lung adenocarcinoma (32.6%) are the top three most KRAS mutant primary cancer types. The metastatic tumors showed a higher prevalence (90.99% versus 66.31%) and diversity of KRAS mutation compared with the primary tumors. Mutations in codons 12 and 13 are the predominant genetic alteration in KRAS (84.15% for TCGA and 86.13% for MSK-IMPACT). Moreover, KRAS mutation is highly correlated with the overall survival of patients with metastatic cancer. The snapback primer assay showed a more favorable performance in enriching and detecting the KRAS codon 12 and 13 mutation (1% mutation load) compared with Sanger sequencing (20% mutation load and 7% false-negative rate). KRAS mutation pattern is highly diverse among different cancer types and is associated with the survival of patients with metastatic cancers. The snapback primer assay is a reliable, sensitive method to detect the major mutant KRAS alleles, which might facilitate the effective cancer treatment decisions.
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DOI：
10.1093/annonc/mdt205
发表时间：
2013-09
期刊：
Annals of oncology : official journal of the European Society for Medical Oncology
影响因子：
--
作者：
Dearden S;Stevens J;Wu YL;Blowers D
通讯作者：
Blowers D
KRAS 突变是 NSCLC 不良预后和治疗结果的弱但有效的预测因子：41 项研究的荟萃分析
影响因子：
--
作者：
Pan W;Yang Y;Zhu H;Zhang Y;Zhou R;Sun X
通讯作者：
Sun X
DOI：
10.1038/nrm3255
发表时间：
2011-12-22
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
通讯作者：
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DOI：
10.1038/nrc3106
发表时间：
2011-10-13
期刊：
Nature reviews. Cancer
影响因子：
--
作者：
通讯作者：
--
影响因子：
50.3
作者：
Stephen, Andrew G.;Esposito, Dominic;McCormick, Frank
通讯作者：
McCormick, Frank