SDC2和TFPI2基因甲基化检测在结直肠癌早期筛查中的临床意义
Establishment and Evaluation of A Multiplex Fluorescence Quantitative PCR Detection Method for Colorectal Cancer Based on Detecting Methylation of SDC2 and TFIP2 Genes
蔡成喜 1马春光 1孙妍兰 1陶吉兰 1张志芳 1马海滨 1徐铭 2刘辉光3
作者信息
- 1. 青海省西宁市疾病预防控制中心(810007)
- 2. 青海省西宁市城东区疾病预防控制中心(810007)
- 3. 青海省西宁市回族医院(810007)
- 折叠
摘要
目的:建立一种快速、准确的结直肠癌(CRC)多重荧光定量PCR早期筛查方法,为该疾病的早期诊疗提供技术支持.方法:基于SDC2基因和TFPI2基因序列,采用MEGA和Primer软件进行序列比对和设计引物、探针,采用10种不同细胞系对引物、探针的灵敏度和特异度进行评价,并对72例经内镜+组织病理活检确诊患者的临床样本和24例对照组样本进行检测评价.结果:本研究设计引物探针能特异性地识别两种CRC细胞系,其他细胞系未见明显的荧光扩增信号;灵敏度分析显示该方法对两个基因(SDC2和TFPI2)的检出限均为100 μg/mL,不同稀释度检测Ct值差异存在统计学意义(P＜0.05).ROC曲线分析显示两个基因(SDC2和TFPI2)检测结果的曲线下面积分别为0.568和0.589.结论:本研究初步构建了一种CRC早期筛查的多重荧光定量PCR方法,在实验室评价中有较好的特异性和灵敏度,但在临床应用中尚需改良优化.
Abstract
Objective:Establish a rapid and accurate multiplex fluorescent quantitative PCR early screening method for colorectal cancer,providing technical support for the early diagnosis and treatment of the disease.Methods:Based on the SDC2 gene and TFPI2 gene sequences,MEGA and Primer software were used for sequence alignment and design of primers and probes.The sensitivity and specificity of primers and probes were evaluated using 10 different cell lines.Clinical samples from 72 patients diagnosed by endoscopy and histopathological biopsy and 24 control samples were tested and evaluated.Results:The primer and probes designed in this study can specifically identify two colorectal cancer cell lines,and no obvious fluorescence amplification signals were found in other cell lines;sensitivity analysis showed that the detection limit of the two genes(SDC2 and TFPI2)was 100 μg/mL,and the Ct values detected at different dilutions were statistically significant(P＜0.05).ROC curve analysis showed that the areas under the curve of the two genes(SDC2 and TFPI2)were 0.568 and 0.589,respectively.Conclusion:This study has preliminarily developed a multiplex fluorescent quantitative PCR method for early screening of colorectal cancer,which has good specificity and sensitivity in laboratory evaluation,but still needs improvement and optimization in clinical application.
关键词
结直肠癌/早期筛查/多重荧光定量PCR/黏结蛋白聚糖2/组织因子途径抑制因子2Key words
Colorectal cancer/Early screening/Multiplex quantitative real time PCR/SDC2/TFPI2引用本文复制引用
出版年
2024
NETL
NSTL
万方数据