分析布鲁杆菌性脊柱炎术后不同组织布鲁杆菌病理阳性率和多重PCR检测阳性率差异。

2018年11月至2019年12月，首都医科大学附属北京地坛医院骨科收治的22例行手术治疗的布鲁杆菌性脊柱炎患者，其中手术部位为颈椎1例、胸椎2例、胸腰椎3例、腰椎16例，其中19例行胸腰椎后路病灶清除、减压、内固定、植骨融合术，1例行颈椎前路病灶清除、减压、内固定、植骨融合术，2例患者行椎间孔镜下病灶清除术，术中收集病灶区不同组织标本（髓核、软组终板、黄韧带、纤维环等），采用Gimesa染色等进行布鲁杆菌病理学观察，同时对各组织标本行布鲁杆菌多重PCR检测，使用Fisher’s精确检验对布鲁杆菌病理阳性率和多重PCR阳性率进行对比。

Gimesa染色发现病灶区任一部位组织标本中存在布鲁杆菌则认定为病理结果阳性，否则为阴性。多重PCR检测发现病灶区任一部位组织标本中存在任何种属的布鲁杆菌DNA则认定为PCR结果阳性，否则为阴性。22例患者中，多重PCR检测阳性率90.9%（20/22）高于Gimesa染色阳性率[81.8%（18/22）]，但差异无统计学意义（P = 0.664）。73份组织标本中，多重PCR检测阳性率[43.8%（32/73）]高于Gimesa染色阳性率[32.9%（24/73）]，但差异无统计学意义（P = 0.173）。其中，22份髓核的多重PCR检测阳性率[86.4%（20/22）]高于Gimesa染色阳性率[72.7%（16/22）]，但差异无统计学意义（P = 0.240）；21份软骨终板的多重PCR检测阳性率[38.1%（8/21）]高于Gimesa染色阳性率[23.8%（5/21）]，但差异无统计学意义（P = 0.505）；19份黄韧带的多重PCR检测阳性率[10.5%（2/19）]与Gimesa染色阳性率[10.5%（2/19）]相同；11份纤维环的多重PCR检测阳性率[18.2%（2/11）]高于Gimesa染色阳性率[9.1%（1/11）]，但差异无统计学意义（P = 1.000）。但髓核Gimesa染色阳性率和多重PCR检测阳性率高于其他部位组织标本阳性率，差异均具有统计学意义（P均< 0.001）。

多重PCR检测可以作为布鲁杆菌性脊柱炎的有效检测手段，其敏感性和准确率高，尤其适用于术前诊断不清和术后病理结果为阴性的患者。同时布鲁杆菌多存在于髓核部位，术中清理和收集过程中应重点处理。

To compare the positive rates of histopathology and multiple polymerase chain reaction (PCR) detection in different tissue specimens in surgery of brucellosis spondylitis.

From November 2018 to December 2019, 22 patients with brucellosis spondylitis underwent surgical treatment in Beijing Ditan Hospital, Capital Medical University, whose surgical sites were cervical spine (1 case), thoracic spine (2 cases), thoracolumbar spine (3 cases) and lumbar spine (16 cases); and 19 patients underwent posterior thoracolumbar debridement, decompression, internal fixation, bone graft fusion, 1 patient underwent cervical anterior debridement, decompression, internal fixation, bone graft fusion, 2 patients underwent foraminal debridement. Different tissue specimens (nucleus pulposus, soft endplate, ligamentum flavum, annulus fibrosus, etc.) were collected for Gimesa staining during the operation. While multiple PCR was performed on the tissue specimens, and the positive rates of both methods were comparatively analyzed by Fisher’s exact test.

Among the 22 patients, the positive rate of multiplex PCR was 90.9% (20/22), which was higher than Gimesa staining [81.8% (18/22)], but without significant difference (P = 0.664). Among the 73 tissue samples, the positive rate of multiplex PCR was 43.8% (32/73), which was higher than that of Gimesa staining [32.9% (24/73)], but without significant difference (P = 0.173). There were 22 samples of nucleus pulposus, and the positive rate of multiple PCR detection was 86.4% (20/22), which was higher than that of Gimesa staining [72.7% (16/22)], but without significant difference (P = 0.240). Among the 21 cartilage endplates, the positive rate of multiple PCR detection was 38.1% (8/21), which was higher than that of Gimesa staining [23.8% (5/21)], but without significant difference (P = 0.505). Among the 19 pieces of ligamentum flavum, the positive rate of multiple PCR was 10.5% (2/19), which was the same as Gimesa staining (2/19). Among the 11 pieces of annulus fibrosus, the positive rate of multiplex PCR was 18.2% (2/11), which was higher than that of Gimesa staining [9.1% (1/11)], but without significant difference (P = 1.000). However, the positive rates of both Gimesa staining and multiplex PCR from nucleus pulposus were significantly higher than those of other tissue samples, with significant difference (P < 0.001).

Multiplex PCR detection can be used as effective detection method for brucellosis spondylitis, with high sensitivity and accuracy, especially for patients with unclear preoperative diagnosis and negative postoperative pathological observation. Brucella mostly resides in the nucleus pulposus, which should be dealt with in the process of cleaning and collecting during operation.