实时荧光PCR检测宫颈组织TSLC1基因方法的建立和应用

Real-time Fluorescent PCR Gene TSLC1 in Cervical Tissue and Application of Established Methods

目的 建立实时荧光PCR检测宫颈组织TSLC1基因的方法,提高TSLC1基因检测方法的灵敏度和精密度.方法 用Light Cycler（R） R480 PCR仪根据设计好的引物与探针,在一定的反应条件下检测宫颈组织TSLC1基因,采用罗氏Light Cycler（R） R480 PCR专用软件进行数据分析.结果 该方法最低检测量为103 copies/ml,检测范围为107～102 copies/ml,10次总的Ct值CV为1.7%.结论 所建立的实时荧光PCR检测各类宫颈组织,正常组织低度鳞状上皮内病变（LSIL）、高度鳞状上皮内病变（HSIL）、宫颈癌组织TSLC1基因的表达率分 别为100%,72%,56%,23%,可以作为进一步研究TSLC1基因表达在肿瘤早期诊断中的价值的重要工具.

Objective The establishment of real-time fluorescence PCR detection of cervical tissue TSLC1 gene,TSLC1 gene detection method to improve the sensitivity and precision. Methods With the Light Cycler480 PCR instrument seized under the designed primers and probes,under certain conditions TSLC1 gene in cervical tissue using the Roche Light Cycler＇480 PCR-specific software for data analysis. Results This method was the minimum detectable amount of 10% copies/ml,detection range 10^7-10^2 copies/ml, 10 times the Ct value of the total CV 1.7%. Conclusion Established realtime fluorescent PCR detection of the cervical tissue,normal tissue,low grade squamous intraepithelial lesion（LSIL）,high grade squamous intraepithelial lesions（HSIL）,cervical cancer rates of TSLC1 gene expression was 100% ,72%,56% and 23% ,could serve as a further expression of TSLC1 gene in the early diagnosis of cancer an important tool.