实验与检验医学
實驗與檢驗醫學
실험여검험의학
EXPERIMENTAL AND LABORATORY MEDICINE
2015年
1期
23-27
,共5页
薛洋%曾富春%舒骏%丛伟
薛洋%曾富春%舒駿%叢偉
설양%증부춘%서준%총위
福尔马林固定石蜡包埋%RNA抽提%基因突变检测
福爾馬林固定石蠟包埋%RNA抽提%基因突變檢測
복이마림고정석사포매%RNA추제%기인돌변검측
Formalin-fixed paraffin-embedded%RNA extraction%Gene mutation detection
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。
目的:評估國內福爾馬林固定石蠟包埋(formalin-fixed paraffin-embedded,FFPE)樣本抽提的RNA質量及其臨床應用。方法收集FFPE樣本994例,採用RNA分離純化試劑盒提取純化RNA,檢測提取RNA的質量,併檢測RNA樣本中的EML4-ALK和ROS1基因突變狀態,同時採用Sanger測序法驗證使用RNA樣本進行基因突變檢測的準確性。結果從994例FFPE樣本中分離提取的RNA的A260/A280和A260/A230均能達到1.8以上,平均濃度為204.23ng/μl,992例樣本均能進行有效擴增。 RNA樣本的EML4-ALK和ROS1基因融閤檢測結果與Sanger測序結果一緻。結論從FFPE樣本中分離得到的RNA質量良好,適用于後續的基因突變檢測,可為腫瘤靶嚮藥物的選擇提供科學、可靠的依據。
목적:평고국내복이마림고정석사포매(formalin-fixed paraffin-embedded,FFPE)양본추제적RNA질량급기림상응용。방법수집FFPE양본994례,채용RNA분리순화시제합제취순화RNA,검측제취RNA적질량,병검측RNA양본중적EML4-ALK화ROS1기인돌변상태,동시채용Sanger측서법험증사용RNA양본진행기인돌변검측적준학성。결과종994례FFPE양본중분리제취적RNA적A260/A280화A260/A230균능체도1.8이상,평균농도위204.23ng/μl,992례양본균능진행유효확증。 RNA양본적EML4-ALK화ROS1기인융합검측결과여Sanger측서결과일치。결론종FFPE양본중분리득도적RNA질량량호,괄용우후속적기인돌변검측,가위종류파향약물적선택제공과학、가고적의거。
Objective To evaluate the quality and clinical application of RNA extracted from formalin-fixed paraffin-embed-ded 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 cas-es 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 sequenc-ing. Conclusions The RNA extracted from FFPE samples is of high quality and suitable for subsequent gene mutation detection by qRT-PCR, which can provide scientific and reliable evidence for selection of tumor targeting drugs.