Two multiplex assays that simultaneously identify 22 possible mutation sites in the KRAS, BRAF, NRAS and PIK3CA genes.
Two multiplex assays that simultaneously identify 22 possible mutation sites in the KRAS, BRAF, NRAS and PIK3CA genes.
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DOI：
10.1371/journal.pone.0008802
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发表时间：
2010-01-21
期刊：
影响因子：
3.7
通讯作者：
Zwarthoff EC
中科院分区：
文献类型：
--
作者：
Lurkin I;Stoehr R;Hurst CD;van Tilborg AA;Knowles MA;Hartmann A;Zwarthoff EC
最近，一些随机试验表明，当晚期结直肠癌患者的肿瘤在KRAS、BRAF和PIK3CA基因中含有突变时，他们不会从靶向表皮生长因子受体的治疗中获益。我们开发了两种多重检测方法，可同时筛选KRAS、NRAS、BRAF和PIK3CA基因中的22个核苷酸突变。该分析在来自晚期结直肠癌患者的294个肿瘤DNA样本上进行了验证。在这些样本中，通过序列分析鉴定了119个KRAS密码子12和13突变，126个肿瘤是KRAS野生型，294个样本中有49个由于DNA质量差而分析失败。这两种突变检测方法检测到130个KRAS突变，其中3个密码子61突变，此外还有32个PIK3CA、13个BRAF和6个NRAS突变。在19个肿瘤中，发现了KRAS突变和PIK3CA基因突变。一个肿瘤对于PIK3CA和BRAF都是突变的。总之，突变检测确定了161个具有突变的肿瘤，120个是野生型，13个分析失败。经计算，2种突变测定的材料成本比获得相同数据所需的测序成本低8倍。此外，突变测定的劳动强度较低。我们的结论是，两个多重突变检测的性能优于直接测序上级。此外，这些分析更便宜，更容易解释。所述测定也可用于选择具有其他肿瘤类型的患者。
Recently a number of randomized trials have shown that patients with advanced colorectal cancer do not benefit from therapies targeting the epidermal growth factor receptor when their tumors harbor mutations in the KRAS, BRAF and PIK3CA genes. We developed two multiplex assays that simultaneously screen 22 nucleotides in the KRAS, NRAS, BRAF and PIK3CA genes for mutations. The assays were validated on 294 tumor DNA samples from patients with advanced colorectal cancer. In these samples 119 KRAS codon 12 and 13 mutations had been identified by sequence analysis, 126 tumors were wild-type for KRAS and the analysis failed in 49 of the 294 samples due to poor DNA quality. The two mutation assays detected 130 KRAS mutations, among which were 3 codon 61 mutations, and in addition 32 PIK3CA, 13 BRAF and 6 NRAS mutations. In 19 tumors a KRAS mutation was found together with a mutation in the PIK3CA gene. One tumor was mutant for both PIK3CA and BRAF. In summary, the mutations assays identified 161 tumors with a mutation, 120 were wild-type and the analysis failed in 13. The material cost of the 2 mutation assays was calculated to be 8-fold lower than the cost of sequencing required to obtain the same data. In addition, the mutation assays are less labor intensive. We conclude that the performance of the two multiplex mutation assays was superior to direct sequencing. In addition, these assays are cheaper and easier to interpret. The assays may also be of use for selection of patients with other tumor types.
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影响因子：
11.5
作者：
Feng, YZ;Shiozawa, T;Konishi, I
通讯作者：
Konishi, I
影响因子：
1.8
作者：
Hurst CD;Zuiverloon TC;Hafner C;Zwarthoff EC;Knowles MA
通讯作者：
Knowles MA
DOI：
10.1093/jnci/djp280
发表时间：
2009-10-07
期刊：
Journal of the National Cancer Institute
影响因子：
--
作者：
Siena S;Sartore-Bianchi A;Di Nicolantonio F;Balfour J;Bardelli A
通讯作者：
Bardelli A
影响因子：
3.7
作者：
Sartore-Bianchi A;Di Nicolantonio F;Nichelatti M;Molinari F;De Dosso S;Saletti P;Martini M;Cipani T;Marrapese G;Mazzucchelli L;Lamba S;Veronese S;Frattini M;Bardelli A;Siena S
通讯作者：
Siena S
影响因子：
15.9
作者：
Hafner, Christian;van Oers, Johanna M. M.;Hartmann, Arndt
通讯作者：
Hartmann, Arndt