Ingrid Sander, Anne Lotz, Eva Zahradnik, Monika Raulf
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For 20 EDC holders, deployment times of cloths were varied between 7 and 28 days, 36 EDCs were used to test reproducibility, and for 28 EDCs extraction buffers were varied (with or without 0.05% Tween 20, borate, or phosphate buffer). The influence of storage of cloths at room temperature (2-629 days) or extracts at -80°C (7-639 days), and variation of extract storage temperature (-20°C and -80°C) for long time storage (1.5 years) on the outcome of allergen quantification were tested for about 150 EDCs. The allergens on EDC cloths increased proportionally with deployment time, and allergen loads on parallel sampled tissues were significantly correlated (P < 0.0001, Pearson of log-transformed values 0.91-0.99). Extraction without Tween reduced all results (P < 0.0001, -51% DM, -85% Dp, -60% Tp, -99% Fel d 1, -86% Can f 1, -52% Mus m 1), and borate buffer resulted in lower yields of Mus m 1 (-53%), DP (-45%), and Tp (-27%) than phosphate buffer. Storage of cloths at room temperature significantly decreased Can f 1 levels (P < 0.0001, -4.8% loss for every 30 days), whereas storage of extracts at -80°C decreased DM results (P < 0.0001, -1.2% loss for every 30 days). Extracts stored at -20°C gave at mean 12% higher DM results compared to extracts stored at -80°C for 1.5 years. Several mammalian allergens and also DM antigens could be quantified reproducibly on EDCs from indoor environments. Allergen levels on EDC cloths increased proportionally with deployment time in a period of 4 weeks. 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However, especially for allergens few experiments on validation of this method concerning deployment time or storage and extraction procedure have been performed. The aim of study was to optimize the EDC procedure for sampling of allergens in indoor environments. EDCs were placed in households or day-care centers and after extraction, allergens were quantified by six immunoassays detecting mite antigens (Domestic mites DM, Dermatophagoides pteronyssinus Dp, Tyrophagus putrescentiae Tp) or the main allergens from cat (Fel d 1), dog (Can f 1) and mouse (Mus m 1). For 20 EDC holders, deployment times of cloths were varied between 7 and 28 days, 36 EDCs were used to test reproducibility, and for 28 EDCs extraction buffers were varied (with or without 0.05% Tween 20, borate, or phosphate buffer). 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引用次数: 8
摘要
在静电除尘器(EDCs)上取样内毒素、β -葡聚糖或过敏原是一种方便的暴露评估方法。然而,特别是对于过敏原,很少有关于该方法的部署时间或储存和提取过程的验证实验。本研究的目的是优化室内环境中过敏原取样的EDC程序。将EDC放置于家庭或日托中心,提取后,通过检测螨抗原(家螨DM、pteronyssinus Dermatophagoides Dp、Tyrophagus purescentiae Tp)或猫(Fel d 1)、狗(Can f 1)和小鼠(Mus m 1)的6种免疫分析法定量分析致敏原。对20名EDC持者,布放时间在7 ~ 28天之间,36份EDC用于重复性测试。对于28种EDCs,提取缓冲液是不同的(含或不含0.05% Tween 20、硼酸盐或磷酸盐缓冲液)。对约150个EDCs进行了织物常温保存(2-629天)或提取物-80℃保存(7-639天)以及提取物保存温度(-20℃和-80℃)长期保存(1.5年)变化对过敏原定量结果的影响。EDC布上的过敏原数量随放置时间的增加呈比例增加,平行取样组织上的过敏原负荷呈显著相关(P < 0.0001,对数变换后的Pearson值为0.91-0.99)。没有Tween的提取降低了所有结果(P < 0.0001, -51% DM, -85% Dp, -60% Tp, -99% Fel d 1, -86% Can f 1, -52% Mus m1),硼酸盐缓冲液导致Mus m1 (-53%), Dp(-45%)和Tp(-27%)的产量低于磷酸盐缓冲液。织物在室温下储存显著降低了Can f 1水平(P < 0.0001,每30天损失-4.8%),而提取物在-80°C下储存降低了DM结果(P < 0.0001,每30天损失-1.2%)。与在-80°C保存1.5年的提取物相比,在-20°C保存的提取物的DM结果平均高出12%。几种哺乳动物过敏原和DM抗原可在室内环境的EDCs上定量再现。在4周的时间内,EDC布上的过敏原水平随部署时间成比例地增加。过敏原产量受提取程序的强烈影响;建议使用清洁剂Tween 20和磷酸盐缓冲液。
Allergen Quantification by Use of Electrostatic Dust Collectors (EDCs): Influence of Deployment Time, Extraction Buffer, and Storage Conditions on the Results.
Sampling of endotoxin, beta-glucan, or allergens on electrostatic dust collectors (EDCs) is a convenient method for exposure assessment. However, especially for allergens few experiments on validation of this method concerning deployment time or storage and extraction procedure have been performed. The aim of study was to optimize the EDC procedure for sampling of allergens in indoor environments. EDCs were placed in households or day-care centers and after extraction, allergens were quantified by six immunoassays detecting mite antigens (Domestic mites DM, Dermatophagoides pteronyssinus Dp, Tyrophagus putrescentiae Tp) or the main allergens from cat (Fel d 1), dog (Can f 1) and mouse (Mus m 1). For 20 EDC holders, deployment times of cloths were varied between 7 and 28 days, 36 EDCs were used to test reproducibility, and for 28 EDCs extraction buffers were varied (with or without 0.05% Tween 20, borate, or phosphate buffer). The influence of storage of cloths at room temperature (2-629 days) or extracts at -80°C (7-639 days), and variation of extract storage temperature (-20°C and -80°C) for long time storage (1.5 years) on the outcome of allergen quantification were tested for about 150 EDCs. The allergens on EDC cloths increased proportionally with deployment time, and allergen loads on parallel sampled tissues were significantly correlated (P < 0.0001, Pearson of log-transformed values 0.91-0.99). Extraction without Tween reduced all results (P < 0.0001, -51% DM, -85% Dp, -60% Tp, -99% Fel d 1, -86% Can f 1, -52% Mus m 1), and borate buffer resulted in lower yields of Mus m 1 (-53%), DP (-45%), and Tp (-27%) than phosphate buffer. Storage of cloths at room temperature significantly decreased Can f 1 levels (P < 0.0001, -4.8% loss for every 30 days), whereas storage of extracts at -80°C decreased DM results (P < 0.0001, -1.2% loss for every 30 days). Extracts stored at -20°C gave at mean 12% higher DM results compared to extracts stored at -80°C for 1.5 years. Several mammalian allergens and also DM antigens could be quantified reproducibly on EDCs from indoor environments. Allergen levels on EDC cloths increased proportionally with deployment time in a period of 4 weeks. Allergen yields are strongly influenced by the extraction procedure; the use of detergent Tween 20 and phosphate buffer is recommended.