All allergic patients and 21 out of 47 of the enrolled sABD underwent SPT to A. simplex using commercial extracts (Laboratorios LETI, Spain; 165 mcg/mL). SPTs were performed on the volar side of the forearm by using disposable 1-mm-tip lancets and were also conducted with histamine (Roxall; 10 mg/ml) as positive and saline solution (0.9% NaCl) as negative controls. Readings were taken at 20 min after application. A mean wheal diameter of ≥ 3 mm was considered positive. sIgE to whole A. simplex extract (Anisakis-sIgE) was determined by ImmunoCAP system (Thermo Fisher Scientific Inc., Uppsala, Sweden) according to the manufacturer’s instructions. A positive result was considered Anisakis-sIgE > 0.35 kUA/l. The same method was used to determinate total IgE, and sIgE to fish.
Immunocap system
Manufactured by Thermo Fisher Scientific
Sourced in Sweden, United States, Germany, Denmark, United Kingdom
The ImmunoCAP system is a fully automated immunoassay analyzer used for the quantitative measurement of specific IgE antibodies in human serum or plasma. It provides accurate and reliable results for the in vitro diagnosis of allergic diseases.
Automatically generated - may contain errors
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120 protocols using immunocap system
1
Anisakis-specific IgE Determination
2
Determining IgE Levels with Immunoassay
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IgE level determinations were performed using the Immuno-CAP system at Thermo Fisher (Allerød, Denmark) on native and IgE-depleted sera (see above). Determination of specific IgE before and after depletion was performed to confirm that componentspecific IgE was effectively removed by ImmunoCAP treatment without affecting the other component-specific IgEs. To exclude that the ImmunoCAP treatment itself impacted the level of specific IgE, serum depletion with an ECP ImmunoCAP was included as control. Furthermore, an IgE depletion by peanut ImmunoCAP was included to demonstrate that all IgE specificities against peanut were removed (for further details refer to online supplementary material).
3
Serum IgG4 and IgE Determination
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The determination of specific IgG 4 and IgE to DP and DF was performed using the ImmunoCAP system (Thermo Fisher, Waltham, Mass., USA). Serum samples for IgG 4 determination were diluted 1:50 before analysis.
4
Serum IgE Measurement by ImmunoCAP
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Total and allergen-specific (SX-1) IgE concentrations in serum were determined using the ImmunoCAP system (Thermo Fisher Scientific, Uppsala, Sweden). The SX1 screening panel detects 8 allergen-specific IgE antibodies: against house dust mite (Dermatophagoides pteronyssinus), cat, dog, birch, timothy (Phleum pratense), rye, ragweed, Cladosporium herbarum.
5
Measuring Af-specific IgG (Gm3) Levels
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Af-specific IgG (Gm3) concentration was measured in our routine laboratory by the commercial ImmunoCAP system (Phadia, Thermo Fisher Scientific, USA), which uses automated fluorescent enzyme immunoassay (FEIA) technology. The m3 antigen is a crude extract from whole Af conidia and mycelia. According to manufacturer data, the reference value is <39 mg/L.
6
Assessing IgE Sensitization in Pediatric Asthma
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IgE-mediated immune reactions are the most common cause of antigen-induced pediatric asthma. Total IgE and allergen-specific serum IgE, our study inclusion factors (Table 1 ), were assessed using the ImmunoCAP System (Thermo Fisher Scientific/Phadia AB, Uppsala, Sweden). The cut-off level was 60 kU/L. In the asthma group, total IgE was greater than 60 kU/L. IgE Serology atopic sensitization was indicated if the child had allergen–specific serum IgE ≥ 0.35 kU/L. We tested allergen-specific serum IgE on dust mite and food allergens of Fx5 (egg white, milk, fish, wheat, peanut, and soy) (22 (link)).
7
Measuring Dpt and Dfa-specific IgE
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Serum was analyzed for total Dermatophagoides pteronyssinus (Dpt)—specific I and Dermatophagoides farinae (Dfa)—specific IgE by the ImmunoCAP system (Thermo Fisher Scientific, Dreieich, Germany).
8
Quantifying Allergen-Specific IgE in Children
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We measured sIgE levels for cow’s milk extract and egg white on an ImmunoCAP system (UniCAP®, Thermo Fisher Scientific, Waltham, MA, USA) with the appropriate allergen caps according to manufacturer instructions. Milk-specific sIgE values were considered positive if they were >15 kU/L in children aged ≥1 year and 5 kU/L in those aged < 1 year [21 ]. Egg-specific IgE values were considered positive if they were >7 kU/L in children aged ≥2 years and 2 kU/L in those aged <2 years [21 ].
9
Measuring Allergen-Specific IgE Levels
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Allergen-specific immunoglobulin E (IgE) was measured against house dust mite, cat, and timothy grass. Analysis was performed in a single central laboratory (AMC Amsterdam) using the ImmunoCAP system (Thermo Fisher Scientific, Uppsala, Sweden). The assay was calibrated against the WHO standard for IgE with a range of 0.35–100 kU/l. A positive IgE was defined as ≥ 0.35 kU/l for one or more of house dust mite, cat, and timothy grass. In this study, data on IgE sensitisation were available only from 3542 subjects.
10
Serum IgE Levels for Respiratory Allergies
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A serum sample was collected from all participants in order to investigate the total IgE level in their blood, as well as some specific IgEs for respiratory allergies (tree pollen, grass pollen, and house dust mite). Blood samples were collected at the Antwerp University Hospital by a responsible nurse. Total and specific IgEs were quantified by an ImmunoCAP System (Thermo Fisher Scientific, Uppsala, Sweden). All assays were performed and results interpreted according to the manufacturers’ recommendation. Total IgE counts below 114 kU/l were considered as non-allergic. For specific IgE counts, values below 0.35 kUA/l were considered as non-allergic.
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