INDOOR Biotechnologies Inc, Charlottesville, VA 22903, USA.
J Immunol Methods. 2013 Jan 31;387(1-2):89-95. doi: 10.1016/j.jim.2012.09.015. Epub 2012 Oct 22.
Consistent performance of allergen assays is essential to ensure reproducibility of exposure assessments for investigations of asthma and occupational allergic disease. This study evaluated intra- and inter-laboratory reproducibility of a fluorescent multiplex array, which simultaneously measures eight indoor allergens in a single reaction well.
A multi-center study was performed in nine laboratories in the US and Europe to determine the inter-laboratory variability of an 8-plex array for dust mite, cat, dog, rat, mouse and cockroach allergens. Aliquots of 151 dust extract samples were sent to participating centers and analyzed by each laboratory on three separate occasions. Agreement within and between laboratories was calculated by the concordance correlation coefficient (CCC).
Results were obtained for over 32,000 individual allergen measurements. Levels covered a wide range for all allergens from below the lower limit of detection (LLOD = 0.1-9.8 ng/ml) to higher than 6800 ng/ml for all allergens except Mus m 1, which was up to 1700 ng/ml. Results were reproducible within as well as between laboratories. Within laboratories, 94% of CCC were ≥ 0.90, and 80% of intra-laboratory results fell within a 10% coefficient of variance (CV%). Results between laboratories also showed highly significant positive correlations for all allergens (~0.95, p<0.001). Overall means of results were comparable, and inter-laboratory CV% for all allergens except Rat n 1 ranged between 17.6% and 26.6%.
The data indicate that performance criteria for fluorescent multiplex array technology are reproducible within and between laboratories. Multiplex technology provides standardized and consistent allergen measurements that will streamline environmental exposure assessments in allergic disease.
过敏原检测的一致性表现对于确保哮喘和职业性过敏性疾病暴露评估的可重复性至关重要。本研究评估了荧光多重阵列的实验室内和实验室间再现性，该阵列可在单个反应孔中同时测量八种室内过敏原。
在美国和欧洲的九个实验室进行了一项多中心研究，以确定尘螨、猫、狗、鼠、蟑螂过敏原的 8 重阵列的实验室间变异性。将 151 份灰尘提取物样本的等分试样分发给参与中心，并由每个实验室在三个不同的时间点进行分析。通过一致性相关系数（CCC）计算实验室内和实验室间的一致性。
获得了超过 32000 个单独过敏原测量的结果。所有过敏原的水平范围很广，从低于检测下限（LLOQ=0.1-9.8ng/ml）到高于所有过敏原的 6800ng/ml，除了 Mus m 1，其高达 1700ng/ml。结果在实验室内和实验室间都是可重现的。在实验室内部，94%的 CCC 值≥0.90，80%的实验室内部结果在 10%的变异系数（CV%）范围内。实验室间的结果也显示出所有过敏原的高度显著正相关（~0.95，p<0.001）。总体结果平均值具有可比性，除了 Rat n 1 之外，所有过敏原的实验室间 CV%在 17.6%至 26.6%之间。
数据表明，荧光多重阵列技术的性能标准在实验室内部和实验室间都是可重现的。多重技术提供了标准化和一致的过敏原测量，这将简化过敏性疾病的环境暴露评估。
