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使用新开发的免疫测定法检测鱼类加工过程中雾化的鱼类抗原。

Detection of fish antigens aerosolized during fish processing using newly developed immunoassays.

作者信息

Lopata Andreas L, Jeebhay Mohamed F, Reese Gerald, Fernandes Joshua, Swoboda Ines, Robins Thomas G, Lehrer Samuel B

机构信息

Division of Immunology/Allergy Section, Faculty of Health Sciences, NHLS Institute of Infectious Disease and Molecular Medicine, Werner Beith Building, Cape Town, Observatory 7925, South Africa.

出版信息

Int Arch Allergy Immunol. 2005 Sep;138(1):21-8. doi: 10.1159/000087354. Epub 2005 Aug 8.

Abstract

BACKGROUND

Aerosolization of fish proteins during seafood processing has been identified as a potential route for allergic sensitization and occupational asthma among workers involved in high-risk activities. The aim of this study was to develop immunological assays for the quantification of aerosolized fish antigens in a fish-processing factory.

METHODS

Polyclonal antibodies to the main fish species processed in the factory (anchovy and pilchard) were generated in rabbits and compared by ELISA inhibition assay and immunoblotting. These antisera were utilized to develop ELISA assays for the detection of fish antigens. The ELISA inhibition assays were evaluated by analyzing environmental air samples collected from three areas in a fish-processing factory: pilchard canning, fish meal production and lobster processing.

RESULTS

By immunoblotting, the rabbit polyclonal antibodies demonstrated IgG antibody binding patterns comparable with IgE antibodies of fish-sensitized patients, particularly in regard to the major fish allergens parvalbumins. The sensitivity of the fish-specific ELISA assays developed was 0.5 microg/ml. The ELISA inhibition assays were able to differentiate between the two different fish species of interest but did not recognize a crustacean species. Notable differences in exposure levels to canned pilchard and anchovy antigens were demonstrated in the three different working areas of the factory, with assays having a detection limit as low as 105 ng/m(3).

CONCLUSION

These ELISA-based assays are sensitive and specific to quantify differential exposure levels to fish antigens produced during fish processing, making it possible to investigate exposure-disease response relationships among workers in this industry.

摘要

背景

在海鲜加工过程中,鱼蛋白的雾化已被确定为从事高风险活动的工人发生过敏致敏和职业性哮喘的潜在途径。本研究的目的是开发免疫测定法,用于量化鱼类加工厂中雾化鱼抗原的含量。

方法

在兔体内产生针对工厂加工的主要鱼类(凤尾鱼和沙丁鱼)的多克隆抗体,并通过酶联免疫吸附测定抑制试验和免疫印迹法进行比较。利用这些抗血清开发用于检测鱼抗原的酶联免疫吸附测定法。通过分析从鱼类加工厂的三个区域采集的环境空气样本,对酶联免疫吸附测定抑制试验进行评估:沙丁鱼罐头加工、鱼粉生产和龙虾加工。

结果

通过免疫印迹法,兔多克隆抗体显示出与鱼类致敏患者的免疫球蛋白E抗体相当的免疫球蛋白G抗体结合模式,特别是在主要鱼类过敏原小清蛋白方面。所开发的鱼类特异性酶联免疫吸附测定法的灵敏度为0.5微克/毫升。酶联免疫吸附测定抑制试验能够区分两种不同的目标鱼类,但不能识别甲壳类物种。在工厂的三个不同工作区域,罐头沙丁鱼和凤尾鱼抗原的暴露水平存在显著差异,测定法的检测限低至105纳克/立方米。

结论

这些基于酶联免疫吸附测定的方法灵敏且特异,可量化鱼类加工过程中产生的鱼抗原的不同暴露水平,从而有可能研究该行业工人的暴露-疾病反应关系。

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