荧光原位杂交技术检测乳腺癌HER2基因状态的影响因素

Application of fluorescence in situ hybridization in the detection of HER2 genetic state of breast cancer

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 obtain the ideal method for detection of HER2 genetic state of breast cancer by using fluorescence in situ hybridization （FISH）. Methods The tissue samples were taken from30 breast cancer patients the influences of digestion time and reusing of denaturization liquid of three different kind of enzymes in the experiment, by adopting the FISH to detect HER2 genetic state were observed. , Results The results showed that the duration of digestion of 15 minutes was not enough and 300 minutes was over digested then the opptimum time was 21minutes, with which the succesful rate of cancer cell hybridization was greater than 75% and the experiment was stable. Conclusion A duration of digestion of enzymes of 21 is suggested for detection of HER2 genetic starus by using FISH and the the succesful rate would be high.