FISH检测中标本切片预处理的优化方案的探索

摘要

目的 探索荧光标记原位杂交技术(FISH)检测中标本切片预处理的优化方案.方法 收集300例明确诊断为肺结核的患者作为阳性对照组,300例健康志愿者(明确排除肺结核患者)作为阴性对照组.收集800例新收临床疑似肺结核患者作为研究组.收集该800例患者的肺活检标本以及同期的痰液标本,其中痰液进行涂片、结核杆菌分离培养、结核杆菌DNA荧光定量PCR检测以及FISH检测.采用两种石蜡包埋组织切片的预处理方案,方案1为自主设计的病理组织切片预处理方法,方案2为参考常用石蜡包埋组织切片预处理方法而设计的方案,比较两种方案对肺结核诊断阳性符合率和阴性符合率.结果 (1)方案1检测结核杆菌阳性符合率为98.7％,方案2检测结核杆菌阳性符合率为97.9％,两种方法阳性符合率差异无统计学意义(P=0.148).(2)方案1检测结核杆菌阴性符合率为97.8％,方案2检测结核杆菌阴性符合率为96.1％,两种方法阴性符合率差异有统计学意义(P=0.036).结论 采用FISH技术诊断肺结核具有较高的价值;与此同时,探索实验条件的优化能够进一步提高FISH诊断的阳性符合率和阴性符合率.%Objective To investigate the optimal approach for sample preparation in the process of FISH evaluation.Methods 300 patients diagnosed as pulmonary tuberculosis and 300 subjects with ruled-out pulmonary tuberculosis were enrolled and served as positive and negative groups respectively.800 suspected patients of pulmonary tuberculosis were also enrolled as research group and underwent sputum smear,culture for mycobacterium tuberculosis,real-time PCR and FISH evaluation.Two approaches of paraffin embedding for pulmonary tissues were used.The first one was self-created novel method and the second one was as traditional process.The sensitivity and specificity of these two methods were compared.Results (1) The positive rate of the first approach was 98.7％ and the second approach was 97.9％ and there was no statistical significant difference between these two methods (P=0.148).(2) The negative rate of the first approach was 97.8％ and the second was 96.1％ and there was statistical significant difference between these two methods (P=0.036).Conclusion FISH was valuable for the tuberculosis detection and optimizing the process would increase the sensitivity and specificity of FISH evaluation.