比较实时荧光PCR法与Sanger测序法检测甲状腺乳头状癌BRAF基因突变

Comparison of real-time PCR method with Sanger sequencing for detection of BRAF mutation in papillary thyroid carcinoma

目的分析实时荧光PCR(real-time PCR)法与Sanger测序法检测甲状腺乳头状癌患者BRAF V600E基因突变的阳性率及符合率。方法收集312例甲状腺乳头状癌手术切除标本,分别采用real-time PCR法和Sanger测序法检测甲状腺乳头状癌中BRAF V600E基因突变情况。结果 312例甲状腺乳头状癌标本real-time PCR法和Sanger测序法检出BRAF基因突变的阳性率分别为65.4%(204/312)、63.8%(199/312)。BRAF基因突变与患者性别无关,与患者年龄相关;两种方法检测结果总符合率为98.4%。结论 real-time PCR法适用于BRAF基因突变的检测。

Purpose To investigate the positive rate and concordance rate of BRAF mutation in papillary thyroid carcinoma detected by real-time PCR method and Sanger sequencing. Methods 312 papillary thyroid carcinomas patients were enrolled in this study. Real-time PCR method and Sanger sequencing were performed to detect BRAF gene mutations. The frequency of BRAF mutation and the concordance of two methods were analyzed. Results BRAF mutation was detected in 65. 4% （204/312） and 63. 8% （199/312） of 312 papillary thyroid carcinoma samples by using real-time PCR method and Sanger sequencing, respectively. There was no significant correlation between BRAF gene mutations and patients’ gender. There was significant correlation between BRAF gene mutations and patients’ age. The overall concordance between real-time PCR method and Sanger sequencing for BRAF mutation detection was 98. 4%. Conclusion Real-time PCR method provides an effective method in BRAF gene mutation detection.