梅毒免疫印迹法在梅毒血清固定患者诊断中的临床应用

Clinical Application of Syphilis Immunoblotting in Diagnosis of Sero-fast Syphilitic Patients

目的探讨免疫印迹法在梅毒血清固定患者诊断中的应用价值。方法使用美国雅培i2000化学发光微粒子免疫分析仪对2017年6月至2020年1月河北燕达医院收治的57612例患者进行梅毒血清学筛查,选取36例梅毒筛查化学发光微粒子免疫法(CMIA)相对发光单位(RLU)/cut-off RLU(S/CO)值≥1的梅毒血清固定患者样本作为血清固定组,30例S/CO值<1的患者样本作为对照组。血清固定患者根据快速血浆反应素环状卡片试验(RPR)基线滴度检测结果进行分组,采用梅毒血清学梅毒螺旋体免疫印迹法(TP-WB)确认,同时检测TpN15、TpN17、TpN45和TpN47的表达,采用MetaboAnalyst 4.0统计分析平台对数据进行统计分析和可视化。结果以TP-WB结果为参照,不同基线滴度样本TpN15、TpN17、TpN45和TpN47的色带着色强度不同,TpN47显色强度最低,TpN15、TpN17、TpN45对应的条带着色强度较高,且不同程度展宽。TpN15、TpN45抗体表达水平在基线滴度为1∶4的样品中显著高表达。相关分析结果显示,TP-WB法TpN15、TpN17、TpN45、TpN47抗原条带读数与CMIA读数均呈正相关(r=0.510,P=0.001;r=0.540,P=0.001;r=0.600,P<0.001;r=0.410,P=0.010),Biplot评估结果显示,各读数值贡献的大小依次为TpN15>TpN17>TpN45>TpN47>CMIA。不同基线滴度样本TpN15、TpN17、TpN45和TpN47的色带着色强度不同,其中1∶2基线滴度的分组样品TpN15抗体表达水平显著高于1∶8基线滴度的分组样品。结论监测梅毒免疫印迹中抗体表达水平变化评估梅毒血清固定状态,对梅毒血清固定患者的诊断具有重要价值。CMIA、RPR以及TP-WB联合检测可以提高梅毒血清固定患者的诊断价值。

Objective To investigate the clinical application value of immunoblotting in the diagnosis of sero-fast syphilitic patients.Methods A total of 57612 patients in Hebei Yanda Hospital from Jun.2017 to Jan.2020 were screened by chemiluminescent microparticle immunoassay(CMIA)Abbott i2000 analyzer,and 36 syphilis screening samples with CMIA relative light unit(RLU)/cut-off RLU(S/CO)≥1 were selected as a sero-fast group,and 30 cases with S/CO value<1 served were included as a control group.According to the results of rapid plasma reagin ring card test(RPR)baseline titer test,the sero-fast samples were divided into several groups.The samples were confirmed by syphilis serology and Treponema pallidum-Western blotting(TP-WB),the expressions of TpN15,TpN17,TpN45 and TpN47 were detected simultaneously,and MetaboAnalyst 4.0 statistical analysis platform was used for statistic analysis and visulization.Results Using the results of TP-WB as references,the color intensities of TpN15,TpN17,TpN45 and TpN47 were different in different baseline titers.TpN47 had the lowest color intensity,and the corresponding bands of TpN15,TpN17,and TpN45 had higher color intensity and widened to varying degrees.TpN15 and TpN45 antibody expression levels were significantly high in samples with a baseline titer of 1∶4.The results of correlation analysis showed that there was a positive correlation between the readings of TpN15,TpN17,TpN45 and TpN47 band by TP-WB method and the readings of CMIA(r=0.510,P=0.001;r=0.540,P=0.001;r=0.600,P<0.001;r=0.410,P=0.010).Biplot evaluation results showed that the contribution of each reading value was TpN15>TpN17>TpN45>TpN47>CMIA.Different baseline titer samples of TpN15,TpN17,TpN45 and TpN47 had different color band intensities.Among them,the TpN15 antibody expression levelof the 1∶2 baseline titer samples was significantly higher than that of the 1∶8 baseline titer samples.Conclusion The detection of antibody expression level in syphilis immunoblotting is of great value in the diagnosis of sero-fast syphilitic patients.CMIA,RPR and TP-WB combined detection can improve the clinical diagnosis value for sero-fast syphilitic s patients.