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目的:探讨荧光原位杂交(FISH)技术检测乳腺癌HER-2基因的最佳实验方法。方法:应用FISH技术检测55例乳腺癌组织标本的HER-2基因,分别采用水浴法和直接消化法,甲酰胺法和共变性法,检测HER-2基因表达成功率,对比实验方法的最佳结果。结果:两种消化法中,水浴法和直接法杂交成功率分别为86.21%(25/29)和88.46%(23/26),差异无统计学意义(P>0.05),杂信号比分别为17.24%(5/29)和46.15%(12/26),差异有统计学意义(P<0.05)。两种变性法中甲酰胺法和共变性法杂交成功率分别为73.08%(19/26)和93.10%(27/29),差异有统计学意义(P<0.05)。结论:FISH检测乳腺癌HER-2基因,以水浴法和共变性法杂交成功率最高。
Objective: To investigate the best experimental method for detection of HER-2 gene in breast cancer by fluorescence in situ hybridization (FISH). Methods: The HER-2 gene was detected by FISH in 55 specimens of breast cancer. The success rate of HER-2 gene expression was tested by water bath method, direct digestion method, formamide method and co-denaturation method, respectively, and compared with the best experimental method result. Results: In both digestion methods, the success rate of hybridization between water bath and direct method was 86.21% (25/29) and 88.46% (23/26), respectively, with no significant difference (P> 0.05) 17.24% (5/29) and 46.15% (12/26) respectively, the difference was statistically significant (P <0.05). The two crossover methods of formamide and co-denaturation hybridization success rates were 73.08% (19/26) and 93.10% (27/29), the difference was statistically significant (P <0.05). Conclusion: The HER-2 gene of breast cancer detected by FISH has the highest success rate of hybridization with water bath and co-degeneration method.