破伤风抗毒素效价絮状单位测定方法(体外法)的验证及其与体内法的比较

Verification of method for determination of titer of tetanus antitoxin by flocculation(in vtiro)and comparison with in vivo test

目的对破伤风抗毒素絮状单位测定法(体外法)进行方法验证,并与小鼠中和试验法(体内法)进行比较。方法对用于破伤风抗毒素效价检测的絮状单位测定法进行准确度、精密度、专属性及耐用性验证。采用絮状单位测定法及小鼠中和试验法分别检测破伤风抗毒素原料血浆、原液及成品,并分析两种方法的相关性。结果对破伤风抗毒素国家标准品6次絮状测定回收率均为100%;重复性验证的变异系数(CV)均<2%;对破伤风抗毒素效价检测具有良好的专属性,供试品中防腐剂对其检测无干扰;不同温度下检测结果的CV值<2%。两种方法对原料血浆、原液及成品测定结果的相关系数分别为0.825、0.937及0.955,P均<0.001,呈正相关,且相关性显著。结论破伤风抗毒素絮状单位测定法(体外法)操作简便,具有良好的准确性、精密性、专属性及耐用性,且与体内法具有显著相关性,可作为企业内控的一种辅助快速检测手段。

Objective To verify the method for determination of titer of tetanus antitoxin by flocculation（in vitro）methodologically and compare with neutralization test in mice（in vivo）. Methods The flocculation test was verified for accuracy,precision,specificity and durability. The plasma as raw material,bulk and final product of tetanus antitoxin were determined by flocculation test and neutralization test separately,of which the correlation of results was analyzed.Results All the recovery rates of national standard for tetanus antitoxin in six flocculation test were 100%,while the CVs in reproducibility verification were less than 2%. The method showed high specificity in determination of tetanus antitoxin titer,while the stabilizer in test samples showed no interference to the test result. The CV of test results at various temperatures were less than 2%. The correlation coefficients of two methods in determination of material plasma,bulk and final product were 0. 825,0. 937 and 0. 995 respectively（each P〈0. 001）,indicating a positive and significant correlation. Conclusion The flocculation test（in vitro） for tetanus antitoxin titer was simple to handle,which showed high accuracy,precision,specificity and durability as well as significant correlation to the in vivo test,and might used as an auxiliary rapid determination method for internal control of manufacturers.