自制与进口肺炎衣原体单克隆抗体的特性比较  

Comparison of Characteristics between Self-made and Imported Monoclonal Antibody to Chlamydia Pneumoniae

作　　者：王卫群[1] 钱利生[2] 史益军[1] 李雪萍[2] 白永怿[1] 余竹元[1] 

机构地区：[1]复旦大学附属金山医院,上海200540 [2]复旦大学上海医学院病原生物学系微生物学教研室,上海200032

出　　处：《中国临床医学》2007年第5期631-633,共3页Chinese Journal of Clinical Medicine

基　　金：上海市卫生局基金资助项目(2002-024006)

摘　　要：目的:自制肺炎衣原体单克隆抗体(Cpn-McAb)与进口Cpn-McAb的特性比较及临床应用。方法:双向琼脂扩散法鉴定自制Cpn-McAb的效价和IgG亚型,用显色法检测单抗与Cpn-MOMP的免疫反应,然后用异硫氰酸荧光素(FITC)标记纯化的IgG。为了评估Cpn-McAb,用自制和进口Cpn-McAb同时检测外周血标本454例。另外,分别从Cpn特异性抗原(Cpn-Ag)阳性的外周血单核细胞(PBMC)标本(A组)中和Cpn-Ag阴性的对照组(B组)中随机各挑选40例,用PCR检测PBMC中的Cpn-DNA。结果:免疫琼脂扩散结果表明,Cpn单抗为IgG2b亚型,扩散效价为1:128。在1:30工作浓度下,微量免疫荧光检测的结果表明,自制Cpn-McAb与鹦鹉热衣原体有较弱的交叉反应;与沙眼衣原体无交叉反应,特性与进口Cpn-McAb一致。自制Cpn-McAb检测PBMC的Cpn-Ag阳性率为53.3%(242/454),进口Cpn-McAb检测PBMC的阳性率为52.6%(239/454),两种单抗检测同时阳性的标本207例,Kappa值是0.704,根据其判断标准提示两者有较高的一致性。A、B两组Cpn-DNA阳性标本分别为38例和2例。结论:自制Cpn-McAb几乎和进口Cpn-McAb一样有较高的特异性和敏感性,有可能替代进口单抗用于临床Cpn感染的检测,有助于相关疾病的诊治。Objective：To compare the characteristics between self-made monoclonal antibody to Chlamydia pneumoniae（Cpn-McAb） and Cpn-McAb from USA,and clinical application of both Cpn-McAb. Methods：The titer of Cpn-McAb and IgG subclass were identified by two dimensional immunodiffusion test, immunological reaction of the antibody with Cpn-MOMP was tested by chromogenic metod,then purified IgG was labeled with fluorescein isothiacyanate. In order to evalute the Cpn-McAb, the blood specimens of 454 patients were detected by self-made Cpn-McAb and Cpn-McAb from USA at the same time. On the other hand, we randomly selected 40 peripheral blood mononuclear cell （PBMC） specimens with Cpn-andgen positive （group A） and 40 ones with Cpn-antigen negative （group B）, then Cpn-DNA in PBMC was detected by PCR. Results： The titer of 1 ： 128 was identified by two dimensional immunodiffusion test for self-made monoclonal antibody, it belonged to IgG2b class and anti-Cpn-MOMP. In the 1：30 working concentration, the outcome of micro-immunofluorescence showed that it had the weak cross reaction with the C. psittaci elementary body but had no cross reaction with the C. trachoma. It has same speciality of Cpn- McAb from USA. The Cpn antigen positive rates of PBMC specimens were 53.3 % （242/454） for self-made Cpn-McAb and 52. 6%（239/454） for Cpn-McAb from USA, and 207 cases were both positive detected by two McAb, showing the high concordance between them （Kappa = 0. 704）. The specimens of positive Cpn-DNA were in 38 and 2 in group A and group B respectively. Conclusion：The self-made Cpn-McAb has almost the same high specificity and sensitivity as Cpn-McAb from USA. The self- made Cpn-McAb may be replace Cpn-McAb from USA to detect Cpn specific antigen and be helpful to diagnose and treat the clinical diseases associated with Cpn infection.

关 键 词：肺炎衣原体单克隆抗体 微量免疫荧光 外周血单核细胞 

分 类 号：R563.1[医药卫生—呼吸系统]