AC群脑膜炎球菌与b型流感嗜血杆菌联合疫苗多糖含量免疫速率比浊检测方法的建立及验证

Development and verification of immune rate turbidimetric method for quantitative determination of polysaccharide content in groups A and C meningococcus and Haemophilus influenzae type b combined vaccine

目的建立AC群脑膜炎球菌与b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)联合疫苗(Men AC-Hib)多糖含量免疫速率比浊检测方法,并进行验证。方法采用免疫速率比浊法测定联合疫苗中A群、C群脑膜炎球菌及Hib多糖含量,绘制不同多糖抗原含量的标准曲线,并对方法的检测低限、精密度、准确度和特异性进行验证。结果 3种不同多糖抗原浓度在2~8μg/ml之间具有良好的线性关系,相关系数均大于0.98;建立的方法检测A群、C群脑膜炎球菌和Hib多糖的检测低限分别为0.847、0.043和0.507μg/ml;检测各组分多糖抗原含量的批内和批间变异系数均小于5%,平均回收率在88.30~104.40%之间;各组分特异性多糖抗体仅与各自多糖抗原反应,且各组分多糖抗原共同存在时无相互干扰。结论成功建立了准确、特异的AC群脑膜炎球菌与Hib联合疫苗的多糖含量检测方法。

Objective To develop and verify an immune rate turbidimetric method for determination of polysaccharide content in groups A and C meningococcus and Haemophilus influenzae type b（Men AC-Hib）combined vaccine. Methods The contents of polysaccharides of groups A and C meningococcus and Hib in Men AC-Hib combined vaccine were determined by rate immune turbidimetric method, based on which the standard curves of antigen contents of various polysaccharides were plotted, and the developed method was verified for minimum detection limit, precision, accuracy and specificity. Results The standard curves of the three kinds of polysaccharides showed good linearity at antigen concentrations of 2 ~ 8 μg/ml, with a correlation coefficient of more than 0. 98. The minimum detection limits of polysaccharides of groups A and C meningococcus and Hib were 0. 847, 0. 043 and 0. 507 μg/ml respectively. Both the coefficients of variation of antigen contents of various polysaccharides in intra-and inter-assays were less than 5%, with mean recovery rates of 88. 30% ~ 104. 4%. The specific antibody against various polysaccharides showed only reactions with the corresponding polysaccharide antigens, while no interference was observed between various polysaccharide antigens. Conclusion An accurate and specific method for determination of polysaccharide contents of Men AC-Hib combined vaccine was successfully developed.