基于qRT-PCR检测非小细胞肺癌患者血浆miR-205方法的建立及临床应用  

Establishment and clinical application of a qRT-PCR method for detecting plasma miR-205 in patients with non-small cell lung cancer

作　　者：李芳琼[1] 王伟[1] 张捷 孟冠敏[1] 刘琴[1] 赵桂芝[3] 

机构地区：[1]浙江省立同德医院检验科,浙江杭州310012 [2]上海之江生物科技股份公司,上海200000 [3]浙江省立同德医院科教科,浙江杭州310012

出　　处：《中国卫生检验杂志》2016年第6期824-827,831,共5页Chinese Journal of Health Laboratory Technology

基　　金：浙江省公益技术研究社会发展项目(2013C33199);浙江省医药卫生科技计划项目(2014KYB053)

摘　　要：目的建立一种定量检测血浆miR-205的qRT-PCR检测方法,并探讨其在NSCLC早期诊断及预后监测中的临床应用价值。方法针对miR-205的序列,构建qRT-PCR扩增体系。收集27例NSCLC患者术前和术后的血浆以及29例健康人血浆进行miR-205测定,并对受试者行ROC分析。结果本研究建立的qRT-PCR法检测miR-205的灵敏度为1.53×10^-9μmol/L,精密度实验CV值均〈5%。NSCLC患者手术前血浆中miR-205表达水平高于对照组,差异具有统计学意义（P〈0.05）;与术前相比,术后一周血浆中miR-205的表达明显降低,差异有统计学意义（P〈0.05）。miR-205所得ROC曲线下面积为0.765（95%CI：0.63-0.899）,区别NSCLC患者和健康体检者的敏感度为77.8%,特异度为69.2%。结论 NSCLC患者血浆中miR-205的表达情况,可作为NSCLC潜在的生物标志物,本实验建立的血浆miR-205 qRT-PCR检测方法具有良好的精密度和灵敏度,有望成为一种新的有效辅助NSCLC早期诊断及判断预后的方法。Objective To establish a specific quantitative detection method based on real time fluorescent quantitative reverse transcriptase- polymerase chain reaction（ qRT- PCR） for plasma micro RNA 205（ miR- 205）. And to explore its clinical value of the early diagnosis and prognosis of non- small cell lung cancer（ NSCLC）. Methods For the sequence of miR- 205,to construct the qRT- PCR amplification system. 27 cases of NSCLC patients＇ preoperative and postoperative plasma and 27 cases of healthy human plasma were collected for the detection of miR- 205,and the subjects were conducted for ROC analysis. Results The minimum detection limit of qRT- PCR method we established was 1. 53 × 10^-9μmol /L; experimental precision CV values were less than 5%. The plasma miR- 205 expression level of NSCLC patients before surgery was higher than control group,the difference was statistically significant（ P〈0. 05）; miR- 205 expression significantly decreased after the operation compared with its preoperative level,and the difference was statistically significant（ P〈0. 05）. miR- 205 showed a receiver operating characteristic（ ROC） curve area of 0. 765（ 95% CI： 0. 63- 0. 899）,the sensitivity was 77. 8%,and the specificity was 69. 2% in discriminating NSCLC patients from the control subjects. Conclusion The plasma miR- 205 expression maybe used as a potential biomarker for non- small cell lung cancer. The qRT- PCR assay,with good precision and sensitivity,is to be a new effective auxiliary method for the early diagnosis and prognosis of NSCLC patients.

关 键 词：非小细胞肺癌 实时荧光定量逆转录PCR miR-205 

分 类 号：R734.2[医药卫生—肿瘤]