自制与进口肺炎衣原体单克隆抗体的特性比较

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。

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

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-antigen 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 detecued 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.