From the enrolled patients we collected samples from tonsils, nasopharyngeal aspirate and peripheral blood. The tonsillectomy was performed according to clinical routine. Tonsil samples were cut into 3–4 mm pieces and stored in RNAlater RNA stabilization reagent (Qiagen, Hilden, Germany), incubated + 4 °C until next working day and after removal of non-absorbed reagent stored in − 80 °C. Nasopharyngeal aspirates were collected at the beginning of the operation during anesthesia using a standardized procedure [11 (link)]. For the viral analysis, part of the tonsil and nasopharyngeal aspirate (NPS) were stored in dry tubes at − 80 °C [11 (link)]. Serum 25(OH)D measurement and serum IgE measurements for food allergen and aeroallergen screening (Phadiatop Combi®, Phadia, Uppsala, Sweden) were made from peripheral blood samples. Patient or his/her guardian also filled a standard questionnaire concerning their respiratory symptoms 30 days prior the operation and allergic diseases (Additional file 1 : Table S1).
Phadiatop combi
Manufactured by Thermo Fisher Scientific
Sourced in Sweden
The Phadiatop Combi® is a laboratory assay used to detect the presence of specific IgE antibodies in human serum. It is designed to screen for a broad panel of common inhalant allergens in a single test.
Automatically generated - may contain errors
Lab products found in correlation
6 protocols using phadiatop combi
1
Biospecimen Collection for Respiratory Research
2
Tonsil Samples Biobanking and Atopy
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Human tonsil samples were obtained from 110 elective tonsillectomy patients (Table 1 ) from Satakunta Central Hospital, Pori, Finland, from April 2008 to March 2009 and biobanked. Tonsillectomy was done according to clinical indications. Written informed consent was obtained from the study patients and/or their guardians. The ethics committee of Turku University Hospital approved the study. All patients filled a standard questionnaire to obtain information of their allergic diseases and respiratory symptoms. Atopy was defined as positive immunoglobulin E (IgE) antibody (>0.35 kU/L) to any of the following allergens: codfish, cow’s milk, egg, peanut, soybean, wheat, cat, dog, horse, birch, mugwort, timothy, Cladosporium herbarum or Dermatophagoides pteronyssinus (Phadiatop Combi®, Phadia, Uppsala, Sweden). Animal sensitization was defined as positive IgE antibodies to cat, dog, horse or Dermatophagoides pteronyssinus. Birch, mugwort, timothy and Cladosporium herbarum were considered as pollen aeroallergens. The eczema was defined as atopic eczema, if a child was atopic and had typical symptoms that included pruritus, typical morphology and chronicity of atopic eczema (Hanifin and Rajka Diagnostic Criteria for Atopic Dermatitis).
3
Childhood Asthma Diagnostic Protocol
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Childhood asthma refers to repeated attacks of airway obstruction and intermittent symptoms of increased airway responsiveness to triggering factors, such as exercise, allergen exposure, and viral infection.12 , 13 “Wheezing” refers to expiratory breathing difficulty as a result of airway obstruction with a high‐pitched bilateral sound during expiration. The presence of allergic sensitization was defined by having one or more positive values for allergen‐specific IgE (>0.35 kU/L, Phadiatop Combi; Phadia, Uppsala, Sweden). Skin prick tests were performed optionally for additional atopy determination using 7 common allergens: normal saline, histamine (10 mg/mL), dermatophagoides mix, cockroach, cat, alternaria, and grass mix in all centers. In addition, birch and Ambrosia were used in Ghent, Lodz, Erlangen and Turku and Parietaria and olive in Athens. Virus infection refers to a positive virus detection using PCR, irrespective of respiratory symptoms.
4
Assessing Allergic Sensitization and Respiratory Viruses
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The methods for inclusion of study subjects and collection of clinical and laboratory data are shown in Appendix S1 . Self‐reported allergies and other clinical data were asked with standard questionnaire (Table S2 ).13 Allergic sensitization was tested with allergen specific immunoglobulin IgE antibodies to any of the common food‐allergen (codfish, cow's milk, egg, peanut, wheat, soybean) or aeroallergens (cat, dog, horse, birch, mugwort, timothy, Cladosporium herbarum and Dermatophagoides pteronyssinus) with cut‐off level 0.35 kU/L (Phadiatop Combi®, Phadia, Uppsala, Sweden) (Table 1 and Table 2 ). Animal sensitization was defined as positive IgE antibodies to cat, dog, horse or Dermatophagoides pteronyssinus. Birch, mugwort, timothy and Cladosporium herbarum were considered as pollen aeroallergens.14 Respiratory viruses were detected from nasopharyngeal aspirates and intratonsillar samples by using PCRs (including reverse transcription step when applicable) on nucleic acid extracts.13 , 15 , 16
5
Allergen Sensitization and Viral Infections
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Atopy (sensitization) was defined as positive immunoglobulin (Ig) E antibody (≥0.35 kU/L) to any of the following allergens: codfish, cow's milk, egg, peanut, soybean, wheat, cat, dog, horse, birch, mugwort, timothy, Cladosporium herbarum and Dermatophagoides pteronyssinus (Phadiatop Combi®, Phadia, Uppsala, Sweden).
Food‐allergen sensitization was defined as positive IgE antibodies to any of the former 6 allergens.
Aero‐allergen sensitization was defined as positive IgE antibodies to any of the latter 8 allergens.
Highly sensitized was defined as IgE > 10 kU/L to any of the allergens detected.
Allergic symptoms were defined as self‐reported allergic rhinitis, doctor‐diagnosed atopic eczema (asked in the questionnaire, Table
Virus infection was defined by positive virus PCR test from nasopharyngeal aspirate (NPA) or tonsil, and virus coinfection if two or more virus PCR tests were positive.
Tonsillar hypertrophy was defined as enlarged tonsils causing symptoms, for example obstructive sleep disorder, snoring and eating problems.
Recurrent tonsillitis was defined as several pharyngitis in the medical records.
6
Defining Atopy and Wheezing in Patients
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