Gastrointestinal Oncology Division, National Cancer Center Hospital, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan.
Anticancer Res. 2013 May;33(5):2129-34.
Direct sequencing (DS) has often been used for detection of v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations. High-resolution melting analysis (HRMA) is another method to detect mutations by using a light scanner, and is more rapid, lower in cost, and more sensitive than DS. We confirmed correspondence between DS and HRMA for KRAS mutation detection in colorectal cancer (CRC).
From 102 patients with CRC, intended to receive cetuximab treatment at the National Cancer Center Hospital between September 2008 and July 2009, formalin-fixed paraffin-embedded tissues were retrospectively analyzed for KRAS status using HRMA and DS. Cetuximab efficacy was evaluated.
Success rates of analysis by DS and HRMA were 100 out of the 102 patients (98.0%) and 99 out of the 102 patients (97.1%), respectively. The cases which failed by one method were analyzed by the other. KRAS mutant-type was detected by DS in 47 out of 100 samples (47.0%), and by HRMA in 45 out of 99 samples (45.5%). The concordance between the two methods was 94.8%. Forty-six out of the 53 patients with wild-type KRAS, as detected by DS received cetuximab and the response and disease control rates were 19.6% and 63.0%, respectively. With a median follow-up of 8.8 months, the median progression-free survival was 5.6 months and median overall survival was 11.1 months. The efficacy of two discordant cases which received cetuximab showed that the best responses were stable disease and progressive disease in one each, progression-free survival was 2.9 and 1.1 months and overall survival was 5.3 and 1.2 months, respectively.
HRMA is a useful optional method for detection of KRAS mutations in CRC in light of accuracy, cost performance and rapidity.
直接测序（DS）常用于检测 v-Ki-ras2 Kirsten 大鼠肉瘤病毒癌基因同源物（KRAS）突变。高分辨率熔解分析（HRMA）是另一种通过光扫描仪检测突变的方法，比 DS 更快、成本更低、更敏感。我们证实了 DS 和 HRMA 在结直肠癌（CRC）KRAS 突变检测中的一致性。
对 2008 年 9 月至 2009 年 7 月期间在国家癌症中心医院接受西妥昔单抗治疗的 102 例 CRC 患者的福尔马林固定石蜡包埋组织进行回顾性分析，使用 HRMA 和 DS 检测 KRAS 状态。评估了西妥昔单抗的疗效。
DS 和 HRMA 的分析成功率分别为 102 例患者中的 100 例（98.0%）和 99 例患者中的 99 例（97.1%）。一种方法失败的病例用另一种方法进行分析。DS 检测到 100 例样本中的 47 例（47.0%）和 HRMA 检测到 99 例样本中的 45 例（45.5%）为 KRAS 突变型。两种方法的一致性为 94.8%。53 例 KRAS 野生型患者中，46 例接受西妥昔单抗治疗，其反应率和疾病控制率分别为 19.6%和 63.0%。中位随访 8.8 个月时，中位无进展生存期为 5.6 个月，总生存期为 11.1 个月。两种不一致病例接受西妥昔单抗治疗的疗效显示，最佳反应分别为稳定疾病和进展性疾病各 1 例，无进展生存期分别为 2.9 个月和 1.1 个月，总生存期分别为 5.3 个月和 1.2 个月。
HRMA 是一种准确、具有成本效益且快速的 CRC 中 KRAS 突变检测的有用选择方法。
