Department of Microbiology and Parasitology, Faculty of Pharmacy, University of Seville, 41012 Seville, Spain.
Biomedal S.L., 41900 Camas, Spain.
Nutrients. 2023 Mar 31;15(7):1730. doi: 10.3390/nu15071730.
A gluten-free diet (GFD) is currently the only treatment available for patients with celiac disease (CD). However, adherence to a GFD can be challenging because gluten is present in many foods. A lifelong follow-up of patients with CD must be performed to promote adherence to a GFD and to identify the appearance of symptoms and the associated diseases. Therefore, the development of tools to analyze gluten exposure in these patients is important. This study proposes the development of the first automatable ELISA to monitor adherence to a GFD through the quantification of urine gluten immunogenic peptides (u-GIP). Seven healthy volunteers without suspicion of CD and 23 patients with CD were monitored as part of this study to optimize, validate, and apply this assay. Non-interference was found in the urine matrix, and the recovery percentage for spiked samples was 81-101%. The u-GIP was stable for up to 16 days when the samples were stored at different temperatures. Overall, 100% of the patients had detectable u-GIP at diagnosis (range of 0.39-2.14 ng GIP/mL), which reduced to 27% after 12 months on a GFD. Therefore, this highly sensitive immunoassay would allow the analysis of u-GIP from a large battery of samples in clinical laboratories of specialized healthcare centers.
目前，对于乳糜泻（CD）患者，无麸质饮食（GFD）是唯一可用的治疗方法。然而，由于麸质存在于许多食物中，因此坚持 GFD 可能具有挑战性。必须对 CD 患者进行终生随访，以促进他们坚持 GFD，并发现症状和相关疾病的出现。因此，开发用于分析这些患者中麸质暴露的工具非常重要。本研究提出了开发首个自动化 ELISA 来监测通过定量尿液中谷蛋白免疫原性肽（u-GIP）来坚持 GFD。本研究监测了 7 名无 CD 怀疑的健康志愿者和 23 名 CD 患者，以优化、验证和应用该检测方法。在尿液基质中未发现干扰，并且添加样本的回收率为 81-101%。当在不同温度下储存时，样品中 u-GIP 的稳定性最长可达 16 天。总体而言，在诊断时，所有患者的 u-GIP 均可检测到（范围为 0.39-2.14ng GIP/mL），在 GFD 治疗 12 个月后降至 27%。因此，这种高灵敏度的免疫测定法将允许在专门医疗保健中心的临床实验室中分析大量样本的 u-GIP。
