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目的分析荧光原位杂交技术(FISH)在乳腺癌HER2基因状态检测中的影响因素。方法选择30例乳腺癌组织标本采用FISH检测HER2基因状态,分别观察三种不同的酶消化时间及重复使用变性液对实验的影响。结果酶消化15min提示消化时间不足,30min提示消化过度,消化21min癌细胞杂交成功率大于75%,实验稳定;变性液重复使用会导致杂交信号减弱或消失。结论在应用FISH检测HER2基因扩增时每次需新配制变性液,酶消化21min,杂交成功率高,且实验稳定。
Objective To analyze the influencing factors of HER2 gene status by fluorescence in situ hybridization (FISH) in breast cancer. Methods The HER2 gene status was detected by FISH in 30 cases of breast cancer tissue samples. The effects of three different enzyme digestion times and the repeated use of denaturing solution on the experiment were observed. Results Enzyme digestion for 15 minutes showed that the digestion time was insufficient, digestion was over 30 minutes, success rate of hybridization of cancer cells was more than 75% after digestion for 21 minutes, and the experiment was stable. Repetition of degeneration liquid could lead to the decrease or disappearance of hybridization signal. Conclusion In the application of FISH detection of HER2 gene amplification each need new preparation of denaturing solution, enzyme digestion 21min, high success rate of hybridization, and experimental stability.