摘要
目的评估国内福尔马林固定石蜡包埋(formalin-fixed paraffin-embedded,FFPE)样本抽提的RNA质量及其临床应用。方法收集FFPE样本994例,采用RNA分离纯化试剂盒提取纯化RNA,检测提取RNA的质量,并检测RNA样本中的EML4-ALK和ROS1基因突变状态,同时采用Sanger测序法验证使用RNA样本进行基因突变检测的准确性。结果从994例FFPE样本中分离提取的RNA的A260/A280和A260/A230均能达到1.8以上,平均浓度为204.23ng/μl,992例样本均能进行有效扩增。RNA样本的EML4-ALK和ROS1基因融合检测结果与Sanger测序结果一致。结论从FFPE样本中分离得到的RNA质量良好,适用于后续的基因突变检测,可为肿瘤靶向药物的选择提供科学、可靠的依据。
Objective To evaluate the quality and clinical application of RNA extracted from formalin-fixed paraffin-embedded samples. Methods A total of 994 FFPE samples were collected and RNA was extracted. The quality of RNA was evaluated and the RNA was used for detection of the gene mutation status of EML4-ALK and ROS1 gene. Sanger sequencing was used for assessing the accuracy of gene mutation detection by using RNA samples. Results The A260/A280 and A260/A230 of RNA from 994 cases of FFPE samples were not less than 1.8, the average concentration of RNA was 204.23ng/μl and all of 992 RNA samples could be effectively amplified. The results of EML4-ALK and ROS1 gene fusion detection were consistent with that of Sanger sequencing. Conclusions The RNA extracted from FFPE samples is of high quality and suitable for subsequent gene mutation detection by q RT-PCR, which can provide scientific and reliable evidence for selection of tumor targeting drugs.
出处
《实验与检验医学》
CAS
2015年第1期23-27,共5页
Experimental and Laboratory Medicine
