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To determine potential biomarkers from the serum and saliva for specific and sensitive diagnosis of eosinophilic esophagitis (EoE).Participants were children aged up to 18 years undergoing esophagogastroduodenoscopy (EGD) evaluation for suspected EoE, follow-up of known EoE, or other upper gastroenterological issues. Participants were recruited from Sachs Children and Youth Hospital, Stockholm, between February 2019 and October 2021.All participants underwent standard EGD procedure and biopsies as per international guidelines with pathologic-anatomic diagnosis. Saliva and blood were collected prior to anesthesia for the EGD procedure. All participants completed a standardized questionnaire regarding allergy history, asthma, eczema, medication use, and any diet restrictions. Blood analytes included absolute eosinophil count (AEC), eosinophil-derived neurotoxin (EDN), specific immunoglobulin E (sIgE) for milk, egg white, wheat, birch pollen, timothy pollen, specific IgG4 for peanut, milk proteins, egg white, soy, birch pollen, and gliadin from wheat. Saliva analytes included 15-hydroxyeicosatetraenoic acid (15(S)-HETE) and total IgG4 in saliva. Statistical evaluation was performed and receiver operating characteristics analysis allowing for calculation of the area under the curve and sensitivity and specificity for various potential markers for EoE.A total of 105 children were included in the study and divided into 3 groups depending on diagnosis by pathology. Thirty-four participants had active EoE meeting criteria of more than 15 eosinophils per high-power field. Eighteen were in EoE remission from previous diagnosis of EoE. Fifty-three had normal histopathology. Combining the following biomarkers—AEC, EDN, sIgE egg white, and sIgE wheat—with reported symptoms (dysphagia, stomach pain, and nausea/vomiting) resulted in an area under the curve of 0.92, allowing excellent discrimination between the 3 groups. Calculated cutoff values resulted in sensitivity of 88% and specificity of 100% in distinguishing those with EoE from healthy individuals.The proposed novel biomarker panel with the addition of symptoms was effective at discriminating between active EoE and healthy individuals. This may contribute to less invasive testing methods for EoE diagnosis and further surveillance for those with active disease.EGD has been the gold standard in diagnosing and accessing improvement in EoE. This invasive procedure is time consuming and carries risk. Novel methods for diagnosis and monitoring response to treatment are needed for EoE. This article proposes such a method that needs additional study to confirm its potential.