Simultaneous identification of 36 mutations in KRAS codons 61 and 146, BRAF, NRAS, and PIK3CA in a single reaction by multiplex assay kit.
Simultaneous identification of 36 mutations in KRAS codons 61 and 146, BRAF, NRAS, and PIK3CA in a single reaction by multiplex assay kit.
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DOI：
10.1186/1471-2407-13-405
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发表时间：
2013-09-03
期刊：
影响因子：
3.8
通讯作者：
Ohtsu A
中科院分区：
文献类型：
--
作者：
Bando H;Yoshino T;Shinozaki E;Nishina T;Yamazaki K;Yamaguchi K;Yuki S;Kajiura S;Fujii S;Yamanaka T;Tsuchihara K;Ohtsu A
西方的回顾性分析表明，KRAS密码子61和146、BRAF、NRAS和PIK 3CA突变是结直肠癌患者接受西妥昔单抗治疗的阴性预测因素。我们开发了一种新型的多重试剂盒，使用Luminex（xMAP）测定法在单个反应中检测KRAS密码子61和146、BRAF、NRAS和PIK 3CA中的36个突变。收集了接受西妥昔单抗治疗的亚洲结直肠癌患者的肿瘤样本和临床数据。我们使用多重试剂盒和直接测序方法研究了KRAS、BRAF、NRAS和PIK 3CA突变，并评估了2种方法之间的一致性。还根据突变状态评价了客观缓解、无进展生存期（PFS）和总生存期（OS）。总共使用两种方法分析了83份样本中的82份（78份手术切除标本和5份活检标本）。使用50 ng模板DNA进行所有多重测定。两种方法之间的一致率为100%。总体而言，49例（59.8%）患者均为野生型肿瘤，21例（25.6%）患者的肿瘤携带KRAS密码子12或13突变，12例（14.6%）患者的肿瘤携带KRAS密码子61、KRAS密码子146、BRAF、NRAS或PIK 3CA突变。这些患者组的缓解率分别为38.8%、4.8%和0%。这些组的中位PFS分别为6.1个月（95%置信区间（CI）：3.1-9.2）、2.7个月（1.2-4.2）和1.6个月（1.5-1.7）;中位OS分别为13.8个月（9.2-18.4）、8.2个月（5.7-10.7）和6.3个月（1.3-11.3）。所有野生型肿瘤患者与KRAS密码子61、KRAS密码子146、BRAF、NRAS或PIK 3CA突变患者之间的PFS和OS均存在统计学显著差异（PFS：95% CI，0.11-0.44; P < 0.0001; OS：95% CI，0.15-0.61; P < 0.0001）。我们新开发的多重试剂盒是实用和可行的调查范围内的样品类型。此外，在亚洲患者中检测到的KRAS密码子61、KRAS密码子146、BRAF、NRAS或PIK 3CA突变不能预测西妥昔单抗治疗的临床获益，与欧洲研究中获得的结果相似。
Retrospective analyses in the West suggest that mutations in KRAS codons 61 and 146, BRAF, NRAS, and PIK3CA are negative predictive factors for cetuximab treatment in colorectal cancer patients. We developed a novel multiplex kit detecting 36 mutations in KRAS codons 61 and 146, BRAF, NRAS, and PIK3CA using Luminex (xMAP) assay in a single reaction. Tumor samples and clinical data from Asian colorectal cancer patients treated with cetuximab were collected. We investigated KRAS, BRAF, NRAS, and PIK3CA mutations using both the multiplex kit and direct sequencing methods, and evaluated the concordance between the 2 methods. Objective response, progression-free survival (PFS), and overall survival (OS) were also evaluated according to mutational status. In total, 82 of 83 samples (78 surgically resected specimens and 5 biopsy specimens) were analyzed using both methods. All multiplex assays were performed using 50 ng of template DNA. The concordance rate between the methods was 100%. Overall, 49 (59.8%) patients had all wild-type tumors, 21 (25.6%) had tumors harboring KRAS codon 12 or 13 mutations, and 12 (14.6%) had tumors harboring KRAS codon 61, KRAS codon 146, BRAF, NRAS, or PIK3CA mutations. The response rates in these patient groups were 38.8%, 4.8%, and 0%, respectively. Median PFS in these groups was 6.1 months (95% confidence interval (CI): 3.1–9.2), 2.7 months (1.2–4.2), and 1.6 months (1.5–1.7); median OS was 13.8 months (9.2–18.4), 8.2 months (5.7–10.7), and 6.3 months (1.3–11.3), respectively. Statistically significant differences in both PFS and OS were found between patients with all wild-type tumors and those with KRAS codon 61, KRAS codon 146, BRAF, NRAS, or PIK3CA mutations (PFS: 95% CI, 0.11–0.44; P < 0.0001; OS: 95% CI, 0.15–0.61; P < 0.0001). Our newly developed multiplex kit is practical and feasible for investigation of a range of sample types. Moreover, mutations in KRAS codon 61, KRAS codon 146, BRAF, NRAS, or PIK3CA detected in Asian patients were not predictive of clinical benefits from cetuximab treatment, similar to the result obtained in European studies.
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KRAS密码子61、146和BRAF突变预测KRAS密码子12和13野生型转移性结直肠癌中对西妥昔单抗和伊立替康的抗性。
影响因子：
8.8
作者：
通讯作者：
--
影响因子：
3.2
作者：
Itoh, Y;Mizuki, N;Inoko, H
通讯作者：
Inoko, H
影响因子：
2.4
作者：
Ogasawara, Naomi;Bando, Hideaki;Esumi, Hiroyasu
通讯作者：
Esumi, Hiroyasu
影响因子：
45.3
作者：
Douillard, Jean-Yves;Siena, Salvatore;Gansert, Jennifer
通讯作者：
Gansert, Jennifer
影响因子：
3.7
作者：
Lurkin I;Stoehr R;Hurst CD;van Tilborg AA;Knowles MA;Hartmann A;Zwarthoff EC
通讯作者：
Zwarthoff EC