基于TaqMan技术构建软组织肉瘤多种融合基因探针及临床辅助诊断应用分析  

Construction of a variety of fusion gene probes for soft tissue sarcoma based on TaqMan technique and their clinical diagnostic applications

作　　者：陈顺平 吴苑 洪少君 李强 翁剑鸣 邹宗楷 蔡铭智 CHEN Shunping;WU Yuan;HONG Shaojun;LI Qiang;WENG Jianming;ZOU Zongkai;CAI Mingzhi(Department of Pathology,Zhangzhou Hospital,Fujian Medical University,Zhangzhou 363000,China)

机构地区：[1]福建医科大学附属漳州市医院病理科,漳州363000

出　　处：《临床与实验病理学杂志》2024年第10期1045-1051,共7页Chinese Journal of Clinical and Experimental Pathology

基　　金：福建省自然科学基金面上项目(2022J011473)。

摘　　要：目的应用TaqMan技术设计PCR联合探针一次性可检测多种软组织肉瘤主要驱动基因的表达,结合病理诊断和FISH结果探讨该联合探针能否较好辅助临床病理诊断需求。方法基于TaqMan技术设计软组织肉瘤相关32对融合基因联合探针,对福建医科大学附属漳州市医院70例相关融合基因软组织肉瘤患者的病理组织学样本进行一次性融合基因联合检测,并设置30例非相关融合基因其他软组织肉瘤患者的病理组织学样本作为对照;常见肉瘤类型结合FISH探针检测结果进行分析;同时对该联合探针检测性能进行多种方法学评价。结果课题组设计的软组织肉瘤相关融合基因探针检测已确诊的软组织肉瘤,结果显示最高敏感性高达100%;选取隆突性皮肤纤维肉瘤、滑膜肉瘤、黏液性脂肪肉瘤3种类型肿瘤结合FISH技术验证,两种方法检测结果一致性高,差异无统计学意义(P>0.05);选取隆突性皮肤纤维肉瘤、滑膜肉瘤、黏液性脂肪肉瘤进行重复性实验,结果完全一致;对所有融合基因RNA阳性质粒进行3种不同浓度检测限检测,确定25拷贝/μL为最低浓度检测下限。结论结合已知病理诊断结果,应用TaqMan技术设计针对软组织肉瘤的PCR联合探针,具备高敏感性和高特异性,方法学性能良好,该探针设计相关融合基因的覆盖位点能满足基层医疗机构一次性快速辅助常见软组织肉瘤病理诊断的需求,为临床软组织肉瘤多种融合基因检测提供了一种快速可靠的检测方法。Purpose To design PCR combined probes using TaqMan technology to detect the expression of major driver genes in a variety of soft tissue sarcomas at one time,and to discuss whether the combined probes can better assist clinicopathological diagnosis based on histological features and FISH results.Methods Our research group designed 32 pairs of fusion gene probes related to soft tissue sarcoma based on TaqMan technique,involving 10 types of sarcoma.The histopathological specimens of 70 patients with common fusion gene soft tissue sarcoma in our hospital were examined by fusion gene combination,and the histopathological specimens of 30 patients with other soft tissue sarcoma without fusion gene were set as controls.Individual common sarcoma types were analyzed with FISH probe detection.At the same time,the detection performance of the combined probe was evaluated by various methods.Results The soft tissue sarcoma-related fusion gene probe designed by our research group was used to detect the confirmed soft tissue sarcomas,and the results showed that the highest sensitivity was 100%.Among the three types of tumors,protuberant dermatofibrosarcoma,synovial sarcoma and mucinous liposarcoma were verified by FISH,and the coincidence rate of the two methods was high,with no statistical significance(P>0.05).The results of interlot and intralot reproducibility of protuberous dermatofibrosarcoma,mucinous liposarcoma and synovial sarcoma were consistent.Three different concentration limits were used to detect the positive plasmid of all the fused gene RNA,and 25 copies/μL was the lowest concentration limit.Conclusion Combined with the pathological diagnosis results,TaqMan technology can be used to design PCR combined probes for soft tissue sarcoma,which have high sensitivity and high specificity and good methodological performance,and meet the needs of primary medical institutions for one-time and rapid auxiliary pathological diagnosis of common soft tissue sarcoma.It provides a rapid and reliable method for the detection of

关 键 词：软组织肉瘤 融合基因 FISH PCR 

分 类 号：R738.6[医药卫生—肿瘤]