Çocuklarda Çoklu Sistemik İnflamatuvar Sendrom (MIS-C) Tanı ve Takibinde Elastin Türevi Peptidlerin Kullanımı ve Tayini İçin Yeni Bir Yarışmalı İndirekt ELISA Protokolü Geliştirilmesi

Abstract

Multisystem Inflammatory Syndrome in Children (MIS-C) is a hyperinflammatory syndrome involving multiple organs, particularly striking cardiac involvement following SARS-CoV-2 infection. Diagnosis of MIS-C relies on clinical assessment, as there is no disease-specific biomarker defined in the literature. Given that elastin is a crucial component of cardiovascular tissue, elastin-derived peptides (EDP) hold promises as a potential biomarker for the diagnosis and prognosis of MIS-C, which is marked by cardiovascular involvement. Therefore, our first objective was to develop a novel ELISA protocol that is practical, rapid, and highly sensitive for the detection of EDP. A novel competitive ELISA protocol was optimized and validated to measure the level of EDP. Secondly, we investigated the role of EDP as a biomarker not only for the diagnosis but also for the prognosis of the cases with MIS-C. A total of 90 children diagnosed with MIS-C included in the patient group, while 65 children included in the control group that is divided two subgroups such as 9 children with severe pneumonia due to SARS-CoV-2 and 56 healthy children.The clinical relevance and usage of EDP as a biomarker candidate, measured by the new competitive ELISA protocol, were investigated in conjunction with the clinical characteristics and laboratory values of the participants. Of the 90 patients with MIS-C, 36 (40%) were characterized by shock, 32 (36%) by a Kawasaki disease-like phenotype and 21 (23%) by a fever & inflammation phenotype. Cardiac involvement was observed in 71 (79%) patients as evidenced by electrocardiographic or echocardiographic or lab findings. Ferritin, CRP, and procalcitonin levels were significantly higher in patients with the shock phenotype compared to those with others. The newly developed competitive ELISA protocol, optimized and evaluated for reproducibility, demonstrated sufficient precision in detecting and quantifying EDP, with an intra-assay coefficient of variation (CV) of 1.96% and an inter-assay CV of 7.64%. EDP levels measured using this novel ELISA protocol in pre-treatment serum samples of patients with MIS-C were significantly lower compared to post-treatment samples, as well as compared to children with SARS-CoV-2 pneumonia and healthy controls. Receiver operating characteristic (ROC) analysis revealed an area under the curve (AUC) of 87%. This study introduces a new and highly sensitive competitive ELISA protocol, paving the way for the utilization of EDP as a novel biomarker in the diagnosis and prognosis of MIS-C.