Detection of Metabolite Biomarkers for the Early Diagnosis and Prognosis of Cow's Milk Allergy in Children
Detection of Metabolite Biomarkers for the Early Diagnosis and Prognosis of Cow's Milk Allergy in Children
In this study, fecal and urine samples will be collected from children diagnosed with :

 -  IgE mediated cow's milk allergy,

 -  suspected of a cow's milk allergy, but with negative diagnosis

 -  IgE mediated food allergy other than cow's milk

 -  healthy brothers and sisters of the first three groups A subset of patients with
 IgE-mediated cow's milk allergy will be asked to provide a urine and fecal sample yearly
 for prognostic purposes.

 The samples will be analyzed using a technique called metabolomics to identify biomarker
 candidates with diagnostic and/or prognostic potential. Additionally, microbiome analysis
 will be performed to map the microbiome of all groups.
In this study urine and fecal samples will be collected from children until the age of 4
 years old. The metabolome and microbiome of children with IgE mediated cow's milk allergy (1)
 (CMA) will be compared to that of children in three control groups: (2) children suspected of
 IgE mediated CMA, but with negative diagnosis, (3) children with an IgE mediated food allergy
 other than cow's milk and (4) healthy brothers and sisters of the first 3 groups. Children
 will be divided into these four groups based on the current diagnostic tests, including IgE
 measurements (skin prick tests and IgE measurements in venous blood samples), clinical
 history of the patients and their response to elimination diets. Samples will be collected
 shortly after diagnosis.

 In a second phase of this study several patients will be followed up for several years (3 - 4
 years), for which yearly a urine and fecal sample will be collected. Children included in
 this part will be younger than 1 year at the moment of their first sample donation, and
 diagnosed with IgE mediated CMA. This longitudinal follow-up is based on the microbial
 difference at the age between 3 to 6 months between children that will have a persistent
 allergy and the ones that will grow out of their allergy. These differences are anticipated
 to be translated in to the metabolomes of these children.

 The samples will be collected at home and have to be frozen immediately after sample
 collection, after which they will be picked up by the researchers. The samples will be
 analyzed using a technique called metabolomics, using UHPLC-HRMS (Ultra-High Performance
 Liquid Chromatography coupled to High-Resolution Mass Spectrometry). After analysis the
 metabolic profiles of the different groups will be compared to each other using multivariate
 statistical analysis. This methodology will be applied to find biomarker candidates with good
 sensitivity and specificity for future CMA diagnosis or prognosis. Additionally, the
 identified biomarkers could provide more insights into the mechanisms behind the disease,
 which can aid in future therapy.

 Microbiome analysis will be performed using qPCR and 16S rRNA sequencing.
Inclusion Criteria:

 -  IgE mediated cow's milk allergy

 -  suspected for a cow's milk allergy, but with negative diagnosis (e.g. non-IgE mediated
 cow's milk allergy)

 -  IgE mediated food allergy other than cow's milk

 -  healthy brothers and sisters from the first three groups

 Exclusion Criteria:

 -  children of 5 years or older