DTP—ELISA试剂盒的试制及应用

Study on Application of ELISA and Its kits For The Detection of Human Serum DTP-Antidodies

本文用 ELISA 法测定了普通人群和免疫前后人血清中白喉、破伤风及百日咳(DTP)抗体水平,并与经典的破伤风抗毒素小鼠中和试验(TNT)、白喉抗毒素家兔皮内中和试验(DNT)及百日咳细菌凝集试验(PA)进行了比较。同时试制出 DTP 血清抗体定性和定量检测药盒。结果表明,在人血清抗体测定中 ELISA 与 TNT、DNT 及 PA 的符合率分别为91.5%、97.9%和91.5%,三者间有良好一致性。20份免疫马血清破伤风抗毒素测定表明 ELISA 与 TNT 相关良好(r=0.84,P<0.01)。以国际上公认的免疫保护水平(TAT≥0.01IU/ml、DAT≥0.01IU/ml 和 PA≥1∶320)为标准,换算成 ELISAP/N 比值(≥2.1)作为试剂盒判断标准,测定了100份普通人群和81例 DTP 接种前后小儿血清,结果100份普通人群血清中含的抗白喉、破伤风及百日咳抗体者分别为28%、38%和45%;接种后小儿血清阳性率分别为88.9%、(白喉)、90.1%(破伤风)和81.5%(百日咳)。ELISA 法具有良好重复性和敏感性,简便经济,可作为临床和流行病学调查及菌苗效果监测的有效工具。

An Enzyme-Linked Immunosorbent Assay (ELISA)has been employed for themeasurement of diphtheria-tetanus-petussis (DTP) antibodies (Mainly IgG) in healthyhuman sera vaccinated with DTP vaccine.In comparison with the in vivo tests (inmice and rabbit) and Bacterial Agglutination (BA),an ELISA kit for the detection ofDTP antibodies was evaluated.Results indicated that there is good agreement betweenELISA and the classical methods in human sera.The correspondences were 91.5% betweELISA and Serum Neutranization Test (SNT);97.9% between ELISA and RabbitSkin Test (RST);91.5%between ELISA and BA,respectively.The correlation of ELISA and SNT in 20 horse sera vaccinated with tetanus toxoid was also good (r=0.84,p<0.01).Converting the criteria of the international commonly considered minimum protective level(i.e.SNT≥0.01 IU/ml;RST≥0.01 IU/ml and BA≥1:320) into P/N ratio (P/N≥2.1)is often employed as a standard for use in the kits,100 human sera and 81 children seravaccinatnated with DTP vaccine were investigated.It was found that protective level ofadult sera was 28% for diphtheria antibody,38% for tetanus antibody and 45% for pertussispertussis antibody respectively.The positive rates of child sera were 88.9% (D),90% (T)and 81.5% (P) respectively.It was concluded that ELISA,having advantages over the classical methods,high re-producibucibility and snd sensitivity,may be used as an acceptable alternativ for theestimatation of DTP antibodies in sero-epidemiological assay,potency surveys of vaccineand clinical purposes with reliablity equal or superior to that of the in vivo tests.