目的 优化现行标准中用于测定转移因子制剂活力的脱E受体法,并建立可用于替代的新方法,用上述方法对来自3个生产企业的57批转移因子制剂进行测定。方法 采用改进的细胞涂片-染色法制备样品的细胞涂片;采用新鲜猪胸腺、兔胸腺分别与绵羊红细胞进行交叉试验;建立微量板-白细胞黏附抑制替代方法,对方法的线性、重复性、专属性等进行考察。结果 改进的涂片-染色法,镜检效果清晰可靠,便于读数和保存。猪胸腺淋巴细胞与兔胸腺淋巴细胞均能与绵羊红细胞形成E玫瑰花结,两种方法试验结果无显著差异。白细胞黏附抑制率结果与玫瑰花结活力测定结果无显著性差异,可用于替代。结论 以改进的染色涂片方法替代原有的细胞计数板法,能更清晰的反应样品的活力值,且可长期保存,适合对大批量样品的随时读片和复核。微量板-白细胞黏附试验替代方法,降低了人为主观因素,较原方法更加准确、快速、重现性好、专属性强,且适合大批量样本的高通量检测。
Abstract
OBJECTIVE To optimize the current T cell-E Rose test method, establish a new alternative method, and measure 57 batches of transfer factor preparations using the two methods. METHODS Cell smear was prepared by modified cell smear-dry staining method. Cross-test was performed with fresh porcine thymus and rabbit thymus respectively with sheep erythrocytes. The micro plate-leukocyte adherence inhibition alternative method was established and verified for the linearity, repeatability, and specificity. RESULTS The improved smear-dyeing method had clear and reliable microscopic effect, which was easy to read the preservation. Porcine thymus lymphocytes and rabbit thymus lymphocytes were able to form E rosette with sheep red blood cells, and there was no significant difference between the two methods. The analysis RESULTS of 57 batches of transfer factor injection showed that the leukocyte adherence inhibition rate had no significant difference with the RESULTS of rosette viability. CONCLUSION The improved method can replace the original cell counting plate method, which can better respond to the actual value of the sample with long-term preservation, and it is suitable for reading and reviewing the analysis result of large quantities of samples at any time. The alternative micro plate-leukocyte adherence test method may reduce the human subjective factors, and is more accurate, rapid, reproducible, and specific than the original method, and suitable for high-throughput analysis.
关键词
转移因子 /
活力测定 /
白细胞黏附抑制 /
质量标准 /
多组分生化药
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Key words
transfer factor /
vitality test /
leukocyte adherence inhibition test /
quality standard /
multi-component biochemical
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中图分类号：
R917
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参考文献
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脚注
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基金
国家自然科学基金项目资助(81603196)
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