检测23价肺炎球菌多糖疫苗各型多糖含量的速率散射比浊法的建立

Determination of different types of polysaccharides in 23-valent pneumococcal polysaccharide vaccine by rate nephelometry

建立准确检测23价肺炎球菌多糖疫苗（23-valent pneumococcal polysaccharide vaccine，PPV23）各型多糖（polysaccharide，PS）含量的速率散射比浊法（rate nephelometry，RN）。方法 制备23价PS参考品和抗血清，建立检测PPV23各型PS含量的RN，并对建立的RN进行线性、精密度、准确度和特异性验证。采用建立的RN在质量控制范围内进行应用性研究。结果 在设定的条件下，当PS参考品质量浓度为1.0 -5.0 μg/ml时，PS浓度与相应响应值呈良好的线性关系，决定系数〉0.99。该法的精密度和准确度良好，批内和批间变异系数分别为0.50% - 4.98%和2.81% - 9.52%，加样回收率为90.3% - 118.8%。该法具有良好的特异性，肺炎球菌抗血清仅与同型PS发生特异性反应，不与其他型PS发生交叉反应。应用性研究显示，在质量控制范围内，该法可准确检测放置不同时间的PPV23各型PS含量。结论 建立的RN可有效控制PPV23的内在质量，可用于PPV23各型PS含量检测。

Objective To develope rate nephelometry （RN） for accurate determination of different types of polysaccharides （PSs） in 23-valent pneumococcal polysaccharide vaccine （PPV23）. Methods The 23-valent PS references and anti-sera were prepared. The RN for determination of different types of PSs in PPV23 was developed. The linearity, precision, accuracy, and specificity of the developed RN were validated. The application research was carried out using the RN within the quality control range. Results Under the designed condition, when PS concentrations were in a range of 1.0-5.0 μg/ml, a good linear correlation was shown between PS concentration and its corresponding instrumental response value with coefficient of determination〉0.99. The method had good precision and accuracy, the coefficients of variation of intra- and inter-assay were 0.50%-4.98% and 2.81%-9.52%, respectively, and the adding recovery rates were 90.3%-118.8%. The specificity of the method was good as the pneumococcal anti-sera only reacted with homotype PS, not heterotype PSs. The application research showed that the RN could accurately detect different types of PSs in PPV23. Conclusion The developed RN can effectively control the internal quality of PPV23, and can be applied to determination of different types of PSs in PPV23.