变性梅毒重组抗原免疫印迹技术的建立及其在鉴别梅毒血清学假阳性实验的应用  

Build of Immunoblotting Technique Using Denatured Recombinant Antigen of T.pallidum and Application to Identify False Positive Serum Test Results of Syphilis

作　　者：彭振亚 张银辉 陈安宁 肖桂初 靳晶 邹钢 陆学东[1] PENG Zhen-ya;ZHANG Yin-hui;CHEN An-ning;XIAO Gui-chu;JIN Jing;ZOU Gang;LU Xue-dong(the Eighth Affiliated Hospital of Sun Yat-Sen University,Guangdong Shenzhen 518033,China)

机构地区：[1]中山大学附属第八医院,广东深圳518033

出　　处：《现代检验医学杂志》2023年第2期150-154,共5页Journal of Modern Laboratory Medicine

摘　　要：目的为了评估临床梅毒抗体筛查中出现假阳性实验结果的真实性,建立了变性梅毒重组抗原免疫印迹技术,并应用于对临床非高危人群梅毒抗体筛查阳性血清进行验证确认,分析出现假阳性的可能性。方法采用梅毒TP47,TP45,TP17和TP15重组抗原,将变性后的抗原与非变性的抗原配对包被在硝酸纤维膜载体上,建立梅毒免疫印迹抗体谱检测方法。收集临床经电化学发光免疫分析仪及配套试剂检测的梅毒螺旋体抗体假阳性的非性病高危人群门诊和住院患者血清样本56例、临床确认梅毒患者血清样本25例和健康体检人员样本30例进行方法学验证。并用该方法对1例特殊的“梅毒患者”进行了鉴别。结果根据建立的变性与非变性抗原配对梅毒免疫印迹抗体谱检测结果判定模式,56例临床梅毒螺旋体抗体筛查假阳性样本,对四种变性重组抗原均不发生反应,无阳性条带。但是对二种以上非变性抗原发生弱阳性反应,出现阳性条带;25例梅毒患者对照样本,对四种非变性和变性重组抗原均有反应,出现强阳性条带;30例健康体检人员样本,对四种非变性和变性重组抗原均无反应。梅毒螺旋体特异性抗体阳性特殊“梅毒患者”血清样本,对四种变性重组抗原均为阴性反应,但对四种非变性重组抗原出现阳性反应。患者样本经血清免疫固定电泳分析,结合临床资料诊断为IgGλ型单克隆免疫球蛋白血症。结论变性梅毒重组抗原免疫印迹抗体谱检测,可有效鉴别临床梅毒抗体假阳性样本。但是,对于梅毒患者的确认一定要将检验结果结合患者的流行病学史、临床表现等进行诊断。Objective To evaluate the correctness of the false-positive results of the clinical syphilis serum screening test,the denatured recombinant antigen of T.pallidum(TP)immunoblotting technique was established and it was applied to verify the positive results of the treponemal antibody screening test in the clinical non-highrisk population and analyze the likelihood of false positives.Methods The recombinant antigens TP47,TP45,TP17 and TP15 of T.pallidum were used,and the denatured antigens and non-denatured antigens were paired and coated on nitrocellulose membrane(NC).The immunoblotting antibody spectrum detection method for syphilis was established.56 cases of false positive samples for T.pallidum antibody detected by electrochemiluminescence immunoassay and supporting reagents from outpatient and inpatient patients with clinical non-sexually transmitted diseases,25 samples of confirmed syphilis patients and 30 samples of healthy physical examinees were collected for methodological verification.A special case of syphilis was identified by this method.Results According to the established judgment model of the detection results of denatured and non-denatured antigens paired with western blot antibody spectrum of syphilis,56 clinical samples of T.pallidum antibody screening were false positive,but none of the four denatured recombinant antigens had any reaction and no positive band.However,there were weak positive reactions to more than two non-denatured antigens,and positive bands appeared.The control samples of 25 syphilis patients all reacted to four non-denatured and denatured recombinant antigens of T.pallidum and showed strong positive bands.The samples of 30 health check-ups showed no reaction to four non-denatured and denatured recombinant antigens.The serum samples of special“syphilis patients”with positive specific antibody against Treponema were negative for four denatured recombinant antigens,but positive for four non-denatured recombinant antigens.The patient was diagnosed as IgGλmonoclonal immunogl

关 键 词：梅毒 梅毒螺旋体 重组抗原 免疫印迹技术 生物学假阳性 

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]