免疫印迹法检测TPN15,TPN17,TPN45和TPN47抗体表达在梅毒诊断中的应用评估  

Evaluation of the Application of Immunoblotting Method for the Detection of TPN15, TPN17, TPN45 and TPN47 Antibodies in the Diagnosis of Syphilis

作　　者：王欣俞 崔凯 赵晋文 丁晓娜 王鑫琪 魏殿军 张延海 高德禄 刘朋 郭奕阳[3] WANG Xin-yu;CUI-Kai;ZHAO Jin-wen;DING Xiao-na;WANG Xin-qi;WEI Dian-jun;ZHANG Yan-hai;GAO De-lu;LIU Peng;GUO Yi-yang(Department of Clinical Laboratory,Hebei Yanda Hospital,Hebei Langfang 066251,China;Department of Clinical Laboratory,Hebei Yanda Medical Research Institute,Hebei Langfang 066251,China;Department of Clinical Laboratory,the Fourth Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北燕达医院检验科,河北廊坊066251 [2]河北燕达医学研究院,河北廊坊066251 [3]河北医科大学第四医院检验科,石家庄050000

出　　处：《现代检验医学杂志》2019年第6期82-85,89,共5页Journal of Modern Laboratory Medicine

基　　金：河北省卫生青年科技课题项目(20180899)

摘　　要：目的通过对4种血清学梅毒实验方法比对,评估免疫印迹法检测TPN15,TPN17,TPN45和TPN47抗体表达在梅毒诊断中的应用。方法于2018年6月~2019年6月,收集河北燕达医院28 417例梅毒筛查样本,综合化学发光微粒子免疫(CMIA)检测结果 S/CO值、梅毒螺旋体明胶凝集试验(TPPA)结果及快速血浆反应素环状卡片试验(RPR)检测结果进行分组,确认实验以梅毒螺旋体抗体免疫印迹试验(TP-WB)平行测定3组标本。相关分析采用Pearson r距离法;数据分类可视化采用主成分分析法。采用Medcalc软件进行ROC曲线统计分析,计算曲线下面积,确定CMIA法对梅毒诊断效能最大时的S/CO最佳截断点;不同诊断方法计数资料的比较采用卡方检验。结果以TP-WB结果为参照,ROC曲线分析发现,当CMIA法S/CO截断值为6.79时,敏感度为79.41%,特异度为96.05%,此时曲线下面积最大为0.911,即6.79为CMIA法的最佳诊断截断值。TPN15,TPN17,TPN45及TPN47抗体表达水平与S/CO比值均呈正相关(P<0.05),相关系数r分别为0.87,0.81,0.87及0.76。当标本中免疫印迹法检测TPN15,TPN17,TPN45和TPN47抗体均阳性且着色强度数值高反应性(50~170)时,四种检测方法阳性符合率、总符合率为100%,显示评价方法和比对方法具有高度一致性(P<0.01)。结论活动性梅毒患者TPN15,TPN17,TPN45和TPN47抗体表达均阳性且着色强度高反应性,梅毒抗体单阳梅毒特异性抗体TPN17和TPN47抗体表达减少且抗体着色强度降低,阳性率显著减少(P<0.05)。梅毒抗体结果可疑患者中TPN15,TPN17,TPN45和TPN47抗体表达明显减少,梅毒血清CMIA法筛查S/CO在6.79以下时,建议进一步采用梅毒免疫印迹检测法进行确证。Objective To evaluate the application of immunoblotting method through the detection of TPN15, TPN17, TPN45 and TPN47 expression in the diagnosis of syphilis by comparing with other four serological methods. Methods From June 2018 to June 2019, 28 417 syphilis screening samples were collected from Yanda Hospital of Hebei province. Samples were grouped into three groups according to the results of CMIA, TPPA and RPR.Pearson r distance method was used for correlation analysis, and principal component analysis(PCA) was used for data classification visualization.The ROC curve was generated by Medcalc software, and the area under the curve was calculated to determine the optimal cut-off point of S/CO when the CMIA method was most effective in the diagnosis of syphilis.Chi-square test was used to compare the counting data of different diagnostic methods. Results Taking the TP-WB method as a reference, ROC curve analysis found that when the S/CO cut-off value of CMIA method was 6.79, the sensitivity was 79.41% and the specificity was 96.05%. At this time, the maximum area under the curve was 0.911, that was, 6.79 was the best cut-off value of CMIA method. The antibody expression levels of TPN15, TPN17, TPN45 and TPN47 were positively correlated with the S/CO ratio(P<0.05), and the correlation coefficients r were 0.87, 0.81, 0.87 and 0.76, respectively. Samples with higher TPN15, TPN17, TPN45 and TPN47 antibodies expression as staining intensity values 50~170 were all syphilis positive, the positive coincidence rate and total coincidence rate of the four detection methods were 100%, indicating that the evaluation method and comparison method were highly consistent(P<0.01).Conclusion The expression of TPN15, TPN17, TPN45 and TPN47 antibodies in active syphilis patients were all positive and the staining intensity was highly reactive. The expression of TPN15, TPN17, TPN45 and TPN47 antibodies in patients with suspicious syphilis antibody results were significantly reduced. When the S/CO value of syphilis serum obtained by

关 键 词：梅毒螺旋体 免疫印迹法 TPN15 TPN17 TPN45 TPN47 评估 

分 类 号：R759.1[医药卫生—皮肤病学与性病学] R392.11[医药卫生—临床医学]