| 梁平1 赵锦荣1 惠延平2 孙建斌1 王伟1 汪钦1 包晗1 刘永兰1 金天博3
郭晏海1 张菊1 颜真1.PIK3CA 基因突变的MassARRAY 分子量阵列分析[J].,2011,11(10):1877-1880 |
| PIK3CA 基因突变的MassARRAY 分子量阵列分析 |
| Genotyping of PIK3CA on Gastric Cancer byMassARRAY |
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| DOI： |
| 中文关键词: 胃癌 基因 基因分型 基因突变 |
| 英文关键词: Gastric cancer Gene Genotyping Gene mutation |
| 基金项目:国家重点基础研究发展规划（973）课题资助（2010CB933900），陕西省科技攻关项目（2008K09-09）
和西安市科技计划项目（SF09027）资助 |
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| 中文摘要: |
| 目的：利用MassARRAY 分子量阵列分析系统检测胃癌组织PIK3CA 基因突变。方法：从胃癌石蜡包埋组织中提取基因组，
PCR 反应扩增目的基因片段，MassARRAY 分子量阵列分析系统检测PIK3CA 基因突变；焦磷酸测序验证检测结果。结果：中国西
部地区144 例胃癌组织样本中PIK3CA_E542K(1624G＞A)突变携带率为77.6%，PIK3CA_E545K(1633G＞A)突变携带率为84%。
MassARRAY 分子量阵列分析系统检测结果与焦磷酸测序结果达到100%吻合。结论：建立了MassARRAY 分子量阵列分析系统
检测基因突变的方法，初步建立了中国西北地区汉族人群胃癌组织PIK3CA 基因PIK3CA_E542K(1624G＞A)和PIK3CA_E545K
(1633G＞A)位点突变频数。 |
| 英文摘要: |
| Objective: To investigate the PIK3CA gene mutation by MALDI-TOF mass spectrometry method (MassARRAY).
Methods: Genomic DNA was extracted from paraffin embedding gastric cancer tissues. Targeted PIK3CA gene fragments were amplified
by PCR. The remaining and nonincorporated dNTPs were removed from amplification products by Shrimp alkaline phosphatase(SAP).
Mass spectrometry detection was carried out after primer extension reaction and primer extension reaction cleanup. Results: In 144 paraffin
embedding gastric cancer tissues were detected, the mutation ratio of PIK3CA_E542K(1624G＞A) was 77.6%, and the mutation ratio
of PIK3CA_E545K (1633G＞A) was 84%. All PIK3CA mutation results were the same as pyrosequencing results. Conclusion: The
MAL- DI-TOF mass spectrometry method for PIK3CA mutation detection was successfully established. The mutation frequencies of
PIK3CA_E542K (1624G＞A) and PIK3CA_E545K (1633G＞A) of gastric cancer have primarily been set up and this founding is only
con- cerning with people of Han nationality among nor-thwest China. |
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