Zhao Qing, Chen Ying-Ying, Hou Chen, Zeng Shao-Xue, Zhang Yu-Hui, Zhang Jun-Jun
Department of Ophthalmology, West China Hospital, Sichuan University, Chengdu 610041, China.
Department of Evidence-Based Medicine, West China School of Medicine, Sichuan University, Chengdu 610041, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2020 Jul;51(4):521-525. doi: 10.12182/20200760503.
To quantitatively detect /11 mutations in uveal melanoma (UM) by droplet digital PCR (ddPCR).
Formaldehyde-fixed paraffin-embedded (FFPE) tumor samples were taken from 78 UM patients with enucleation in West China Hospital between 2009 and 2015. None of the patients received radiotherapy or chemotherapy before enucleation. A retrospective study was conducted to detect /11 mutation in UM by ddPCR. To compare the consistency of the results of the two detection methods, DNA sequencing was performed on the target gene by Sanger sequencing. 78 patients with UM were studied retrospectively. /11 mutations in uveal melanoma was detected by ddPCR. The consistency of the results of the two detection methods was analyzed.
/11 mutations frequency was 91.9%. The consistency test between Sanger sequencing and ddPCR of /11 mutations in 74 patients with UM was conducted. coefficient=0.436, =0.001. The error rate of Sanger sequencing results was significantly higher in the heterogeneous group than in the homogeneous group (12/37 vs. 3/16, =0.53), but the difference was not statistically significant.
The results of ddPCR and Sanger sequencing showed good consistency, and the mutation ratio of /11 in UM was significantly different. /11 mutation frequency in UM patients detected by ddPCR was close to the reported frequency. It is more recommended to use ddPCR with high sensitivity to detect gene mutations in samples of tumor tissue DNA derived from FFPE. Sanger sequencing is prone to false negative results.
通过液滴数字PCR（ddPCR）定量检测葡萄膜黑色素瘤（UM）中的/11突变。
收集2009年至2015年期间在华西医院接受眼球摘除术的78例UM患者的甲醛固定石蜡包埋（FFPE）肿瘤样本。所有患者在眼球摘除术前均未接受放疗或化疗。采用回顾性研究，通过ddPCR检测UM中的/11突变。为比较两种检测方法结果的一致性，采用Sanger测序对目标基因进行DNA测序。对78例UM患者进行回顾性研究。通过ddPCR检测葡萄膜黑色素瘤中的/11突变。分析两种检测方法结果的一致性。
/11突变频率为91.9%。对74例UM患者的/11突变进行Sanger测序与ddPCR的一致性检验。系数=0.436，=0.001。异质性组中Sanger测序结果的错误率显著高于同质性组（12/37对3/16，=0.53），但差异无统计学意义。
ddPCR与Sanger测序结果显示出良好的一致性，且UM中/11的突变率存在显著差异。ddPCR检测UM患者的/11突变频率接近报道频率。更推荐使用灵敏度高的ddPCR检测FFPE来源的肿瘤组织DNA样本中的基因突变。Sanger测序容易出现假阴性结果。
