Warthin-Starry特殊染色、免疫组织化学和透射电镜在猫抓病病理诊断中的作用
Application of Warthin-Starry stain,immunohistochemistry and transmission electron microscopy in diagnosis of cat scratch disease
摘要目的 探讨Warthin-Starry银染色法、抗汉赛巴尔通体单克隆抗体和电镜在检测人巴尔通体感染、确诊猫抓病中的实用价值.方法 收集1998年1月至2008年12月诊断的77例符合猫抓病形态学改变的石蜡包埋淋巴结组织,分别使用Warthin-Starry银染色法和抗汉赛巴尔通体单克隆抗体的EnVision法进行病原体的检测.选择二者均检出丰富细菌的5例进行透射电镜观察.结果 (1)透射电镜查见汉赛巴尔通体呈多形性,圆形、椭圆形、短棒状、杆状,长0.489~1.110μm,宽0.333~0.534 tun,集群性生长.(2)61.0%(47/77)的病例Warthin-Starry银染色法查见排列成线、成簇、局灶性聚集的黑色、短小、棒状杆菌,细菌绝大多数分布在靠近被膜下的坏死灶内,以存在于细胞外为主.(3)72.7%(56/77)的病例抗汉赛巴尔通体单克隆抗体阳性表达,阳性信号主要呈点状、颗粒状,少数呈线样勾勒出细菌形状,信号定位与Warthin-Starry银染色法一致.(4)59.7%(46/77)的病例Warthin-Starry银染色和抗汉赛巴尔通体单克隆抗体染色均阳性.两种方法联用,病原体总体检出率为74.0%(57/77).二种方法检出率的差异具有统计学意义(P<0.05).结论 应用Warthin-Starry银染色法、免疫组织化学和电镜技术检测猫抓病病原体能达到确诊猫抓病的目的 ,抗汉赛巴尔通体单克隆抗体染色是较Warthin-Starry银染色更理想的检测方法.
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abstractsObjective To evaluate the diagnostic utility of Warthin-Starry silver stain,immunohiatochemistry and transmission electron microscopy in the detection of human Bartonella henselae infection and pathologic diagnosis of cat scratch disease(CSD).Metbotis The paraffin-embedded lymph node tissues of 77 histologically.defined cases of cat scratch disease collected during the period from January,1998 to December,2008 were retrieved and studied using Warthin-Starry silver stain(WS stain)and mouse monoclonal antibody against Bartonella henselae(BhmAB stain).Five cases rich in bacteria were selected for transmission electron microscopy.Results Under electron microscope,the organisms Bartonella henselae appeared polymorphic,round,elliptical,short rod or bacilliform shapes,ranged from 0.489 to 1.110 μm by 0.333 to 0.534 μm and often clustered together.Black short rod-shaped bacilli arranged in chains or clumps were demonstrated in 61.0%(47/77)of CSD by WS stain.The organisms were located outside the cells and lie mainly in the necrotic debris,especially near the nodal capsule.In 72.7%(56/77)of the cases,dot-like,granular as well as few linear positive signals were observed using BhmAB immunostain and showed similar localization.Positive results for both stains were identified in 59.7%(46/77)of the cases.When applying both stains together,Bartonella henselae was observed in 74.0%(57/77)of the earle.The difference between the results obtained by WS stain and BhmAB immunostain was of statistical significance(P<0.05).Conclusions Bartonella henselae is the causative pathogen of cat scratch disease.WS stain,BhmAB immunostain and transmission electron microscopy are helpful in confirming the histologic diagnosis.Immunostaining using BhmAB can be a better alternative than WS stain in demonstrating the organisms.
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