酶联免疫破伤风抗体定量试剂的研制及应用

Study and application of quantitative ELISA kit in detection of antibody to tetanus

制备一种精确的破伤风抗体定量试剂,用于人源破伤风抗体的定量及人群破伤风抗体水平的测定.以人源破伤风免疫球蛋白国家标准品建立定量反应曲线,经系统优化后建立双抗原夹心法定量检测系统.定量反应曲线显示,抗体浓度在10～120mIU/ml之间,相关系数r = 0.9993.精密度（CV）≤7 %.实际应用验证,未经（TTC）免疫的献血员中,具有保护性抗体水平（0.01IU/ml）的比例只占12.2%.经3针（TTC）免疫后,抗体水平均大于0.01IU/ml,经动物体内中和试验法（NT）定量的3批破伤风免疫球蛋白,用制备的酶联免疫破伤风抗体定量试剂测定,其回收率分别为109%、98%和93%.结论,该试剂可用于破伤风抗体的精确定量.

To develop an accurate quantitative EHSA kit that can be used for the quantifying human antibody to tetanus and estimating antibody responses in tetanus toxoid immunized groups, Linear regression curves were developed by using the national standard preparation （tetanus immunoglobulin from human）. A sandwich ELISA kit was developed by optimizing all kinds of conditions and applicability was validated. The optimal linear range of regression curve was in a range of 10mIU/ ml-120mIU/ml （ r =0. 9993 ）. Precision was equal to or less than 7%. The samples from immunized group were detected by EIA, the result indicated that the ratio for possessing protective antibody only accounted for 12.2% in all tested samples, and after immunized three times, the antibodies all reachod to a protective level above 0.01IU/ml. Three lots of human tetanus immunoglobulins were quantified by neutralization test （ NT）, the repeatabilities were 109%, 98%, 93%, respectively performed by using the ELISA kit.