弓形虫抗血清 IgG 的制备及其检测效果研究  

The evaluation and preparation of antiserum IgG of Toxoplasma

作　　者：楼涤[1] 陈睿[1] 张乐[1] 陆绍红[1] 林爱芬[1] 卓敏敏[1] 傅翠娥[1] 

机构地区：[1]浙江省医学科学院寄生虫病研究所,杭州310013

出　　处：《热带病与寄生虫学》2004年第1期22-24,共3页Journal of Tropical Diseases and Parasitology

摘　　要：目的比较弓形虫不同免疫原所制备抗血清 IgG 的检测效果,优选高效价的抗体诊断试剂。方法制备弓形虫的细胞质抗原(C)、代谢抗原(S)。用抗原 C、C+S、S 及活虫(P)各免疫2只家兔,收集抗血清经纯化而获得 IgG 抗体(多抗)并制备酶标记抗体;用上述4种多抗与对应的多抗-HRP 组合成4种双抗体夹心型 ELISA,检测人血清 CAg 和 C、S 抗原,评价试剂的敏感性;抗血清与疟原虫、隐孢子虫等抗原进行交叉反应性试验,评价试剂的特异性;采用捕获法检测人血清 IgM 抗体,比较4种多抗-HRP 的检测效果。结果各种 ELISA 夹心法同步检测 CAg 阳性标本7例,阴性样本8例,均未出现假阴性和假阳性反应,其 OD 均值的 P/N 比值依次为 C+C 组33.5、S+S 组27.8、CS+CS 组21.2、P+P 组13.2;C 和 S 抗原的最低检出量均为0.5μg/ml;与其它寄生虫未出现交叉反应。4种多抗-HRP 同步检测IgM 抗体阳性标本3例,阴性样本7例,均未出现假阴性和假阳性反应,其 OD 均值的 P/N 值最高的是抗 C-HRP。结论 4种抗体用于检测人血清 IgM 抗体、CAg、C 和 S 抗原以及其它抗原的结果表明,各种抗体试剂均具有良好的敏感性和特异性,其中以抗 C 抗体试剂最为满意。(2)4种组合形式的夹心ELISA,检测弓形虫抗原的差异无统计学意义,表明弓形虫 C 和 S 抗原间存在着共同抗原成分。Objective To compare the effects of antibodies by different Toxoplasma immunogens on detection of CAg,and to select antibody with high titer as diagnostic reagents of Toxoplasma.Methods Cyto- plasmic(C)and metabolic(S)antigen of Toxoplasma were prepared and two rabbits were respectively immu- nized with C,C+S,S antigen and living Toxoplsma(P).To evaluate the sensitivity of reagents,CAg in hu- man sera and C,S antigen were detected by 4 kinds of double-antibody sandwich ELISA composed of 4 polyclonal antibodies and IgG-HRP.Cross-reactions of antisera and Malaria,Cryptosporidium antigen were detected.Results No false positive and false negative results were found by detecting seven CAg positive samples and eight CAg negative samples at the same time.P/N ratio of medium OD value was 33.5,27.8, 21.2 and 13.2 for the group of C+C,S+S,CS+CS and P+P respectively.The least detectable amount of C and S antigen was both 0.5μg/ml.No cross-reactions with other parasites were found.No false positive and false negative results were found by detecting three IgM positive samples and seven IgM negative samples with IgG-HRP at the same time.The highest P/N ratio of medium OD value was anti-cytoplasmic antibody.Con- dusion(1)The results of four antibodies detecting CAg in human sera,C,S and other antigens show that an- tibody reagents above are all highly sensitive and specified,especially the anti-cytoplasmic antibody.(2)No statistic significance was found when Toxoplasma antigen was detected by 4 kinds of composed-sandwich ELISA which indicated that common antigenic components might exist between C and S antigen.

关 键 词：血清IgG 制备 效果研究 双抗体夹心型ELISA ELISA夹心法 IgM抗体 夹心ELISA 假阳性反应 抗体诊断试剂 检测效果 阳性标本 同步检测 酶标记抗体 IgG抗体 血清CAg 交叉反应性 最低检出量 弓形虫抗原 S抗原 隐孢子虫 P/N值 

分 类 号：R446.11[医药卫生—诊断学] R395.6[医药卫生—临床医学]