肺炎衣原体抗体的制备及应用_0.docVIP

肺炎衣原体抗体的制备及应用 作者：王卫群 张瑞华 余竹元 【摘要】 目的 肺炎衣原体抗体(Cpn-PcAb)的制备及 应用 。 方法 用10μl Cpn菌株TW-183(2×108ifu)与300μl福氏完全佐剂混匀后皮下多点注射新西兰白兔。双向琼脂扩散法鉴定血清抗体的效价为164，用硫酸铵盐析法、DEAE-Sephadex-50层析法纯化免疫球蛋白(IgG)，然后用异硫氰酸荧光素(FITG)标记IgG。用Cpn-PcAb与美国进口Cpn-单抗(Cpn-McAb)同时检测病人外周血标本294例。另外，分别从Cpn-PcAb检测出Cpn特异性抗原阳性的外周血单核细胞(PBMC)标本(A组)中和对照组(B组)中随机各挑选20例，用PCR检测PBMC中的Cpn-DNA。结果 Cpn-PcAb检测的阳性率为45.9%(135/294)，Cpn-McAb检测的阳性率为43.5%(128/294)，两种抗体同时阳性的标本有110例，Kappa值是07，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17例和3例。结论 Cpn抗体几乎和Cpn单抗一样有较高的特异性和敏感性，可能可以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。 【关键词】 肺炎衣原体多克隆抗体 肺炎衣原体单克隆抗体 微量免疫荧光 外周血单核细胞 Production and application of polyclonal antibody to chlamydia pneumoniae 【Abstract】 Objective Production and application of polyclonal antibody to chlamydia pneumoniae (Cpn).Methods The New Zealand white rabbit was subcutaneously injected with 10μl Cpn strain TW-183(2×108ifu) and 300μl complete Freund adjuvant.The serum antibody titer of 1∶64 was identified by two dimensional immunodiffusion test.IgG was extracted from serum and purified by ammonium sulphate salting out and DEAE-Sephadex A-50 chromatography，then was labeled by fluorescEin isothiacyanate.In order to evalute Cpn-PcAb，the blood specimens of 294 patients were deected by Cpn-PcAb and Cpn monoclonal antibody (Cpn-PcAb) from USA at the same time.On the other hand，we randomly selected 20 peripheral blood mononuclear cell (PBMC) specimens with Cpn antigen positive (group A) and 20 ones with Cpn antigen negative (group B) by Cpn-PcAb detection，then detected Cpn-DNA in PBMC by PCR.Results The positive rate was 45.9% (135/294) for Cpn-PcAb and 43.5%(128/294) for Cpn-McAb，and 110 cases have the same positive for them，showing the high concordance between them (Kappa=0.7).The specimens of positive Cpn-DNA were in 17 and 3 respectively between group A and group B.Conclusion The Cpn-PcAb has almost the same high specificity and sensitivity as Cpn-McAb.The Cpn-PcAb may be replace Cpn-McAb to detect the Cpn specific antigen and be helpful