Abstract：
Progress towards standardisation of allergen products has been made in recent years. Nevertheless, no standardised test method to quantify the allergen content of grass pollen allergen products is available at present. One aim of the BSP090 project was to validate a quantitative assay for a major Timothy grass (Phleum pratense) pollen allergen, Phl p 5. Qualification of a candidate ELISA system was performed with regard to range, robustness and cross-reactivity in preliminary studies. The assay specifically detected Phl p 5 with a quantification range from 3.9 ng/mL to 62.5 ng/mL. Suitability to quantify recombinant and natural Phl p 5 was further assessed in a collaborative study including 14 laboratories in Europe and the USA. Precision and accuracy of the assay was satisfactory with 93% of calculated Phl p 5 concentrations and 100% of total recoveries being within the ± 30% acceptance range. Similar results were obtained for spike recoveries, with exclusion of the lowest concentration spike, showing spike recoveries exceeding the acceptance range for six laboratories. Inter-assay (repeatability) and inter-laboratory (reproducibility) variability were satisfactory, in the format used in the present study. Robustness towards different statistical methods for data analysis was demonstrated. In conclusion, the assay can easily be established in routine testing and results of the preliminary testing and collaborative study support the proposal of the assessed Phl p 5-specific ELISA as a European Pharmacopoeia general method.
摘要：
近年来在变应原产品的标准化方面取得了进展。然而，目前还没有标准化的测试方法来量化草花粉过敏原产品的过敏原含量。BSP090项目的一个目的是验证主要Timothy草（Phleumpratense）花粉过敏原的定量测定，Phlp5.对候选ELISA系统进行范围鉴定，初步研究中的稳健性和交叉反应性。该测定以3.9ng/mL至62.5ng/mL的定量范围特异性地检测Phlp5。在包括欧洲和美国的14个实验室的合作研究中进一步评估了定量重组和天然Phlp5的适用性。测定的精密度和准确性令人满意，计算的Phlp5浓度为93％，总回收率为100％，在±30％接受范围内。加标回收率也获得了类似的结果，排除最低浓度峰值，显示峰值回收率超过六个实验室的接受范围。分析间（可重复性）和实验室间（再现性）变异性令人满意，在本研究中使用的格式。证明了对不同统计方法进行数据分析的稳健性。总之,该试验可以很容易地在常规检测中建立,初步检测和合作研究的结果支持了作为欧洲药典通用方法的评估的Phlp5特异性ELISA的建议.
