Department of Molecular Genetics and Microbiology, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA.
Department of Internal Medicine, Division of Rheumatology, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA.
Biosens Bioelectron. 2016 Sep 15;83:306-11. doi: 10.1016/j.bios.2016.04.048. Epub 2016 Apr 22.
Antinuclear antibodies (ANA) are important in diagnosis and follow-up of patients with autoimmune conditions. The current increase in ANA requests is driven by broadening the use of ANA from a test for lupus to a test for diverse autoimmune diseases, but the standard method is protracted, cumbersome and prone to error. We describe an electrochemical method for quantifying total ANA for use as a point-of-care diagnostic aid. In this technology the target autoantigens are derived from a protein/nucleoprotein mixture prepared from an inexpensive source and adsorbed to a porous membrane with high protein binding capacity. Serum is slowly drawn through the membrane comprising the high density autoantigen mixture to induce rapid binding of patient autoantibodies. After rinsing, peroxidase-conjugated anti-IgG is drawn through the membrane followed by rinsing, insertion of an electrode assembly, and addition of the enzyme substrate. Substrate peroxidation is measured by microamperage-level current accompanying electrochemical reduction of the intermediate product. Values are comparable to a standard ANA test but require a total processing time of ~20min. This method has the promise to greatly expand ANA testing in clinical settings for initial patient assessment of autoimmune disease.
抗核抗体（ANA）在自身免疫性疾病患者的诊断和随访中非常重要。ANA 请求的当前增加是由于将 ANA 的用途从狼疮检测扩大到了多种自身免疫性疾病的检测，但标准方法冗长、繁琐且容易出错。我们描述了一种用于定量检测总 ANA 的电化学方法，可作为即时诊断辅助工具。在这项技术中，目标自身抗原源自一种从廉价来源制备的蛋白质/核蛋白混合物，并吸附在具有高蛋白质结合能力的多孔膜上。血清缓慢通过包含高密度自身抗原混合物的膜，以诱导患者自身抗体的快速结合。洗涤后，将过氧化物酶缀合的抗 IgG 吸入膜中，然后进行洗涤，插入电极组件，并添加酶底物。通过伴随中间产物电化学还原的微安培级电流测量底物过氧化物酶。该方法与标准 ANA 检测相比具有可比性，但总处理时间约为 20 分钟。这种方法有望极大地扩展临床环境中的 ANA 检测，用于自身免疫性疾病患者的初步评估。
