长臂光敏基因探针对结核性浆膜腔积液的诊断价值

目的 研究聚合酶链反应（PCR）联合长臂光敏生物基因探针对浆膜腔积液性的诊断价值。方法 通过PCR特异扩增人型结核分枝杆菌（H37Rv)IS986基因序列245bp片段，构建含该特异片段的重组质粒，以该重组质粒DNA为模板，用PCR 扩增IS986产物制备长臂光敏生物互标记的基探针，进行探针特异性检测。同时对65份胸腔积液标本进行基因扩增联合 探针检测并与传统的病原学检测方法进行比较。结果 通过PCR联合探针检测结果显示；该探针除了与人型、牛型、田鼠及非洲分枝杆菌有特异杂交信号外，与其他的分枝杆菌没有杂交反应。故此地具有良好的特异性。对65份临床胸腔积液标本进行结核分枝杆菌检测，其阳性检出率（46.66%) 分别高于PCR（26.66%)、涂片（0％）及培养（13.33%)的检出率。结论 该基因探针特异性强、敏感性高，与目前涂片镜检、培养法、单纯PCR扩增相比均有明显优势，适用于临床检测，有推广价值。

The diagnostic value of improved PCR in pleural offusion tuberculosis

Objective To study the diagnotic value of combining PCR with IS986 gene probe marked with long arm light sensitive biotin. Methods The 245 bp fragment of human MTB (H 37 Rv) gene sequence was repeated multiple times through PCR, and established reconstructive plasmid. The gene probe marked with long arm light sensitive biotin was used to detect 65 pleural effusion samples by combining gene amplification with probe examination. Results There was no hybridizing reaction with other Mycobacterium except human MTB, cow MTB, Microtus and Africa Mycobacterium. The probe had significant peculiarity. The results of PCR combined with probe detection showed that its positive rate was higher than that of PCR, smear exam and mycrobacterial culture respectively. Conclusions The gene probe has obvious advantages of stronger peculiarity, higher sensitivity compared with smear examination, mycrobacterial culture and pure PCR. It can be used in clinical examination.