【摘要】 目的尝试用新型自动化原位杂交仪(HS400Pro)建立规范化、标准化操作步骤。方法可有效降低实验所需探针浓度、提高杂交效率。对5例已知HER-2阳性的乳腺癌存档蜡块,采用动态杂交技术行2种不同DNA探针标记(DIG和FITC),分别用手工和HS400Pro全自动杂交检测系统对比检测结果。结果探针用专利杂交液按1∶40稀释后,采用新型自动化原位杂交仪37℃杂交12 h与手工37℃杂交18 h结果一致。说明该仪器可以基本替代手工开展ISH。结论自动化原位杂交仪的使用是实现ISH自动化、标准化、规范化的好方法。但要继续扩大检测样本量,找出HS400Pro的最佳杂交条件。更多还原

【Abstract】 Objective To study the standardization and normalization of HER-2 detection by in situ hybridization.Methods This new automation hybridization station(HS400 Pro automation hybridization system) uses a new agitation hybridization technology,which helps to effectively reduce the concentration of probes and improve hybridization efficiency.Five cases of breast cancer with known Her2 gene amplification were collected in this study.Probes labeled by DIG and fluorescence were selected.Experiments were performed by both manual and automation operation.Results After the probes were diluted with patent hybridization buffer in 1∶40,we used the HS400Pro automation hybridization station to perform hybridization for 12 hs in 37℃,and the results were consistent with manual hybridization for 18 hs in 37℃.It meant this machine was able basically to replace the manual operation of in situ hybridization.Conclusion It is a good way to use this automation hybridization station for automation,standardization and normalization of ISH.Major disadvantage is the few cases studied that is insufficient,and therefore we will further expand the sample size and find the optimal hybridization condition of HS400 Pro.更多还原