Molecular Allergy Research Laboratory, College of Public Health, Medical and Veterinary Sciences, James Cook University, Townsville, Queensland, Australia.
Centre for Food and Allergy Research, Murdoch Children's Research Institute, Melbourne, Victoria, Australia.
Allergy. 2019 Jul;74(7):1352-1363. doi: 10.1111/all.13748. Epub 2019 Apr 15.
Commercial allergen extracts for allergy skin prick testing (SPT) are widely used for diagnosing fish allergy. However, there is currently no regulatory requirement for standardization of protein and allergen content, potentially impacting the diagnostic reliability of SPTs. We therefore sought to analyse commercial fish extracts for the presence and concentration of fish proteins and in vitro IgE reactivity using serum from fish-allergic patients.
Twenty-six commercial fish extracts from five different manufacturers were examined. The protein concentrations were determined, protein compositions analysed by mass spectrometry, followed by SDS-PAGE and subsequent immunoblotting with antibodies detecting 4 fish allergens (parvalbumin, tropomyosin, aldolase and collagen). IgE-reactive proteins were identified using serum from 16 children with confirmed IgE-mediated fish allergy, with focus on cod, tuna and salmon extracts.
The total protein, allergen concentration and IgE reactivity of the commercial extracts varied over 10-fold between different manufacturers and fish species. The major fish allergen parvalbumin was not detected by immunoblotting in 6/26 extracts. In 7/12 extracts, five known fish allergens were detected by mass spectrometry. For cod and tuna, almost 70% of patients demonstrated the strongest IgE reactivity to collagen, tropomyosin, aldolase A or β-enolase but not parvalbumin.
Commercial fish extracts often contain insufficient amounts of important allergens including parvalbumin and collagen, resulting in low IgE reactivity. A comprehensive proteomic approach for the evaluation of SPT extracts for their utility in allergy diagnostics is presented. There is an urgent need for standardized allergen extracts, which will improve the diagnosis and management of fish allergy.
用于过敏皮肤点刺测试 (SPT) 的商业过敏原提取物广泛用于诊断鱼类过敏。然而，目前对蛋白质和过敏原含量的标准化没有监管要求，这可能会影响 SPT 的诊断可靠性。因此，我们试图使用鱼类过敏患者的血清分析商业鱼类提取物中鱼类蛋白的存在和浓度以及体外 IgE 反应性。
检查了来自五个不同制造商的 26 种商业鱼类提取物。测定了蛋白质浓度，通过质谱分析蛋白质组成，然后进行 SDS-PAGE 并随后用检测 4 种鱼类过敏原（副肌球蛋白、肌球蛋白、醛缩酶和胶原蛋白）的抗体进行免疫印迹。使用来自 16 名经证实的 IgE 介导的鱼类过敏儿童的血清鉴定 IgE 反应性蛋白，重点是鳕鱼、金枪鱼和鲑鱼提取物。
不同制造商和鱼类之间，商业提取物的总蛋白、过敏原浓度和 IgE 反应性差异超过 10 倍。免疫印迹法未检测到 6/26 种提取物中的主要鱼类过敏原副肌球蛋白。通过质谱法在 7/12 种提取物中检测到 5 种已知的鱼类过敏原。对于鳕鱼和金枪鱼，几乎 70%的患者对胶原蛋白、肌球蛋白、醛缩酶 A 或β-烯醇酶表现出最强的 IgE 反应性，但不是副肌球蛋白。
商业鱼类提取物通常含有包括副肌球蛋白和胶原蛋白在内的重要过敏原含量不足，导致 IgE 反应性低。提出了一种用于评估 SPT 提取物在过敏诊断中的实用性的综合蛋白质组学方法。标准化过敏原提取物迫在眉睫，这将改善鱼类过敏的诊断和管理。
