| 姜英浩 张潍 俞青苗 强少盈 梁平 程红 高艳娥 赵星烨 张菊.宫颈癌中EGFR启动子甲基化荧光偏振技术检测方法研究[J].现代生物医学进展英文版,2014,14(11):2162-2165. |
| 宫颈癌中EGFR启动子甲基化荧光偏振技术检测方法研究 |
| A Novel EGFR Promoter Methylation Status Assay Based on FluorescencePolarization in Cervical Cancer Tissue Samples |
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| DOI： |
| 中文关键词: 荧光偏振检测 宫颈癌 表皮生长因子受体 甲基化 |
| 英文关键词: Fluorescence polarization Cervical cancer Epithelial growth factor receptor(EGFR) Methylation |
| 基金项目:国家自然科学基金项目（81071435） |
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| 中文摘要: |
| 目的：建立一种基于荧光偏振技术的宫颈癌组织标本中EGFR 启动子甲基化检测的新方法。方法：设计通用引物，在封闭管
中同时扩增EGFR 启动子甲基化与非甲基化等位基因片段，再同时用序列特异的FAM 或TAMRA 荧光标记探针对扩增产物进
行杂交检测，利用荧光偏振仪检测扩增杂交反应的荧光偏振值，确定EGFR启动子甲基化状态。应用荧光偏振法检测63 例宫颈
癌组织样本，并与直接测序法进行对比验证。结果：本方法检测EGFR启动子甲基化结果与直接测序法结果无统计学差异，且最
低检测浓度为50 拷贝/ul，最低检测含量可达10 %，敏感度高。结论：本方法检测EGFR启动子甲基化灵敏度与准确度较高，为临
床宫颈癌个体化治疗相关检测提供了新技术。 |
| 英文摘要: |
| Objective:To develop the novel assay detecting the EGFR promoter methylation status in cervical cancer tissue samples
by using a hybridization-fuorescence polarization(FP) technique. Methods:A pair of primers was used to amplify a 156 bp fragment
in the promoter region of EGFR. Two probes specifc for either methylated or unmethylated EGFR promoter DNA labeled with different
fuorophores hybridized, respectively, with their target amplicons. The EGFR promoter methylation status was determined by the FP values.
A total of 63 cervical cancer tissue samples were analyzed by using the new assay technique and sequencing in parellel. Results:The
results of new assay showed no statistically significant difference with the results of sequencing. The minimumdetection level established
with the new assay was 50 copies/mL, and it was able to detect the minor population of the EGFR promoter methylation status even when
its contents were as low as 10 %. Conclusion:The novel hybridization-FP assay for evaluating the EGFR promoter methylation status
was suitable for the clinical setting and would provide an applicable pharmacogenomic tool for individualized management of cervical
cancer patients. |
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