微流控与重组酶聚合酶扩增用于原肌球蛋白悬浮颗粒物的微量样品采样与分析方法研究

鱼虾蟹贝类水产品加工过程中会产生大量经由空气传播的过敏原悬浮颗粒物。这些过敏原（如原肌球蛋白）颗粒物经呼吸道进入人体后能够引起过敏性哮喘、荨麻疹等症状，严重危害水产加工及水产科研人员的健康。此类有害产物在空气中含量较低且无法像微生物样品那样被培养扩增，因此不适宜直接套用微生物悬浮颗粒物的分析方法，即空气采样器捕获微量样品—微生物培养扩增—微生物分析。.本课题以如何结合运用生物化学分析原理，建立针对性的新方法，实现虾类水产品加工中产生的原肌球蛋白悬浮颗粒物的现场快速分析这一科学问题为主线，提出微流控与重组酶等温扩增相结合的采样与分析方法。该方法具有试剂消耗量少、成本低、设备便携等优势，预计分析时间2小时，检测限达1微克每立方米空气，适用于现场快速分析和工作场所空气质量的评价，进而可监控和预防水产食品加工过程产生的有害产物对人员造成的健康威胁，保证充足的劳动力以维持水产食品行业的可持续发展。

Aerosolization of allergen proteins during the aquatic product processing has been identified as an essential cause for occupational allergy and respiratory disease among workers involved in fishery processing activities. Thus, the pollution of airborne allergen aerosol affects workers’ health in a fishery operation setting and potentially hinders the development of aquatic food industry. As a way to deal with it, the advancement in monitoring technology can greatly improve workplace health and safety service. However, it is still quite difficult to rapidly collect and quantify seafood allergen aerosol owing to its low concentration in air. So far, conventional methods for air sampling and analysis, which have been widely used in the analysis of microorganism densities in aerosols, are not best suited to the analysis of seafood allergen aerosol.. We herein express our serious concern about the personal health risk caused by aerosolized crustaceans allergen such as tropomyosin. The specific aim of this proposal is to establish a novel approach that can achieve the fast collection and accurate quantification of aerosolized tropomyosin particulate which is generated during the processing of shrimps such as Penaeus vannamei Boone and Procambarus clarkii. To this end, the state-of-the-art microfluidics and Recombinase Polymerase Amplification (termed as RPA), which is a nucleic acid isothermal amplification technology, will be employed. In this proposal, the ultimate goal is to develop microfluidics chip assays with 1 μg/m3 limit of detection and 2-hour operation time. The success of our proposed research will make substantial contribution to evaluating air quality and monitoring threat level in a fishery processing setting, and thereby prevent occupational disease and maintain sustainable development of aquatic food industry.