改良的粪便脱落细胞富集方法的富集效率分析

Analysis of enrichment efficiency of the improved fecal exfoliated cells enrichment method

目的分析改良的粪便脱落细胞富集方法的富集效率。方法对收集的102例结直肠癌患者或高危人群的清肠导泻液采用改良富集方法富集脱落细胞,对收集到的细胞悬液提取总DNA和RNA,利用实时定量PCR(quantitative real-time PCR,qRTPCR)法检测标本中内参基因表达(β-actin和RNU6),通过PCR扩增曲线分析富集效率,建立脱落细胞质量控制体系流程。结果 102例样本粪便脱落细胞富集悬液提取总DNA和RNA后内参基因β-actin和RNU6检测信号均为阳性,细胞富集率为100%。结论改良的粪便脱落细胞富集方法的富集效率高,通过分子生物学检测后确定样本可进行后续的标志物验证。对脱落细胞进行分子生物学检测能够简单、快捷地鉴定粪便脱落细胞的质量,为进一步行粪便细胞早诊标志物检测提供基础。

Objective To analyze the enrichment efficiency of improved enrichment method tor tecal extoliated cells. Methods Intestine-rinsing fluid from 102 colorectal cancer patients or high-risk population were made into cell suspension by improved tecal extoliated cells enrichment method. The total DNA and RNA were extracted from the cell suspension and detected the expression of the housekeeping gene （[3-actin and RNU6） by the real-time quantitative PCR （qRT-PCR）. The enrichment efficiency was analyzed by qRT-PCR amplification curve, and the quality control system of extoliated cells was established. Results In 102 tecal extoliated cells suspension samples, the positive expression rates of β-actin gene and RNU6 gene were both 100%. Conclusion The enrichment efficiency of improved tecal extoliated cells enrichment is satisfied. The study supplies a simple and rapid way tot more molecular biology detection of extoliated cells and provides a basis to the explore and verily molecular markers in tecal cells of early colorectal cancer.