# paper.pdf ## Metadata - PDFFormatVersion=1.4 - EncryptFilterName=Standard - IsLinearized=false - IsAcroFormPresent=false - IsXFAPresent=false - IsCollectionPresent=false - IsSignaturesPresent=false - Creator=Unknown - CreationDate=D:20250505120539Z - Author=Unknown - Title=Untitled - Trapped=False - ModDate=D:20250505120539Z - Producer=XEP 4.25.502 ## Contents ### Page 1 ImmunoCAP™ Specific IgEFluoroenzymeimmunoassayCalibrator Range 0-100 kU/l Directions for Use 52-5291-EN/12INTENDED USEImmunoCAP Specific IgE is an in vitro test system for the quantitative measurement of allergen specific IgE in human serum or plasma. It is intended for in vitro diagnostic use as an aid in the clinical diagnosis of IgE mediated allergic disorders in conjunction with other clinical findings, and is to be used in clinical laboratories. ImmunoCAP Specific IgE is to be used with the instruments Phadia 200, Phadia 250, Phadia 1000, Phadia 2500 or Phadia 5000.SUMMARY AND EXPLANATION OF THE TESTAllergic diseases such as rhinitis, conjunctivitis, asthma, atopic dermatitis/eczema, and food allergy represent clinical manifestations of adverse immune responses following repeated contact with usually harmless substances in the environment (1). These substances are often referred to as allergens. Allergic or immediate hypersensitivity reactions may occur within minutes after allergen exposure. The serum factors capable of mediating allergic reactivity in sensitized individuals are antibodies of the immunoglobulin E (IgE) class (2, 3). Atopy is a propensity to produce IgE antibodies in response to normal exposure to common allergens, with or without associated allergy symptoms (1). Sensitization to different allergens can be established by measuring IgE antibodies in blood. Measuring specific IgE antibodies against various allergens is a useful aid to clarify whether observed symptoms are likely to be due to an underlying allergic disease or not (4, 5). Allergen sensitization in predisposed individuals varies depending on the age of the patients, hereditary factors and allergen exposure (6). Also, the symptoms can vary between allergic individuals. It has been shown to be useful to measure and follow the levels of specific IgE antibodies as an aid in the diagnosis of allergic diseases as well as possible tolerance development (4-10). Quantitative measurement of allergen specific IgE antibodies using the ImmunoCAP Specific IgE assay provides an indication of the risk for clinical reactions to an allergen and assists in the identification of offending allergens for avoidance measures (4, 5, 11, 12). A negative test result is a valuable aid to rule out allergy (5). ImmunoCAP Allergens (extract-based ImmunoCAP Allergens and ImmunoCAP Allergen Components) and ImmunoCAP Allergen Mixes can be used as an aid in the clinical diagnosis of patients with suspected IgE mediated allergic disorders. ImmunoCAP Allergen Mixes provide qualitative results of specific IgE to a combination of allergen sources and should be used as a guide for further testing with relevant ImmunoCAP Allergens.PRINCIPLE OF THE PROCEDUREThe allergen of interest, covalently coupled to ImmunoCAP, reacts with the specific IgE in the patient sample. After washing away non-specific IgE, enzyme labeled antibodies against IgE are added to form a complex. Following incubation, unbound enzyme-anti-IgE is washed away and the bound complex is then incubated with a developing agent. After stopping the reaction, the fluorescence of the eluate is measured. The higher the response value, the more specific IgE is present in the sample. To evaluate the test results, the responses for the patient samples are transformed to concentrations with the use of a calibration curve.REAGENTS AND MATERIALReagents are packaged as described below, each purchased separately. The two-digit suffix (-xx) on the article number may vary between countries. All kits are not available in all countries. The expiration date and storage temperature are stated on the labels. Do not use reagents beyond their expiration dates or if damaged or incorrectly stored.Note: It is not recommended to pool any reagents. Keep the ImmunoCAP carrier closed to avoid evaporation of buffer.Reagents for Phadia 200• ImmunoCAP Specific IgE Conjugate 100 (Art No 10-9316-01: for 6 x 100 determinations) • ImmunoCAP Specific IgE Conjugate 400 (Art No 10-9310-01: for 6 x 400 determinations) • ImmunoCAP Specific IgE Calibrator Strip 0-100 (Art No 10-9459-01: for 5 calibration curves) • ImmunoCAP Specific IgE Curve Control Strip (CC-1 and CC-2) (Art No 10-9312-01: 5 x 3 sets of curve control) • ImmunoCAP Specific IgE Anti-IgE (a_IgE) (Art No 14-4417-01: carriers of 16 ImmunoCAP) • ImmunoCAP Allergen and Allergen Mix (See Product catalogue: carriers of 16 or 10 ImmunoCAP) • ImmunoCAP Total IgE Low Range (low) (Art No 14-4497-35: for 48 determinations) • ImmunoCAP Phadiatop (phad) (Art No 14-4405-35: for 48 determinations) • ImmunoCAP Phadiatop Infant (phinf) (Art No 14-4510-35: for 48 determinations) • Development Solution (Art No 10-9441-01: for 6 x 200 determinations; Art No 10-9440-01: for 6 x 315 determinations) • Stop Solution (Art No 10-9442-01: for 6 x 185 determinations; Art No 10-9479-01: for 6 x 100 determinations) • Washing Solution (Art No 10-9422-01: 6 x 1 l) Washing Solution Additive, 6 x 17.2 ml• • Washing Solution Concentrate, 6 x 80 ml • Washing Solution (Art No 10-9202-01: 2 x 5 l) Washing Solution Additive, 2 x 86 ml• • Washing Solution Concentrate, 2 x 400 ml • ImmunoCAP Specific IgE Control L (Art No 10-9528-01: for 6 x 4 determinations) • ImmunoCAP Specific IgE Control M (Art No 10-9529-01: for 6 x 4 determinations) • ImmunoCAP Specific IgE Control H (Art No 10-9530-01: for 6 x 4 determinations) • ImmunoCAP Specific IgE Negative Control (Art No 10-9445-01: for 6 x 4 determinations)Reagents for Phadia 250• ImmunoCAP Specific IgE Conjugate 100 (Art No 10-9316-01: for 6 x 100 determinations) • ImmunoCAP Specific IgE Conjugate 400 (Art No 10-9310-01: for 6 x 400 determinations) • ImmunoCAP Specific IgE Calibrator Strip 0-100 (Art No 10-9459-01: for 5 calibration curves) • ImmunoCAP Specific IgE Curve Control Strip (CC-1 and CC-2) (Art No 10-9312-01: 5 x 3 sets of curve control) • ImmunoCAP Specific IgE Anti-IgE (a_IgE) (Art No 14-4417-01: carriers of 16 ImmunoCAP) • ImmunoCAP Allergen and Allergen Mix (See Product catalogue: carriers of 16 or 10 ImmunoCAP) • ImmunoCAP Total IgE Low Range (low) (Art No 14-4497-35: for 48 determinations) • ImmunoCAP Phadiatop (phad) (Art No 14-4405-35: for 48 determinations) • ImmunoCAP Phadiatop Infant (phinf) (Art No 14-4510-35: for 48 determinations) • Development Solution (Art No 10-9441-01: for 6 x 200 determinations; Art No 10-9440-01: for 6 x 315 determinations) • Stop Solution (Art No 10-9442-01: for 6 x 185 determinations) • Washing Solution (Art No 10-9422-01: 6 x 1 l) Washing Solution Additive, 6 x 17.2 ml• • Washing Solution Concentrate, 6 x 80 ml • Washing Solution (Art No 10-9202-01: 2 x 5 l) Washing Solution Additive, 2 x 86 ml• • Washing Solution Concentrate, 2 x 400 ml • ImmunoCAP Specific IgE Control L (Art No 10-9528-01: for 6 x 4 determinations) • ImmunoCAP Specific IgE Control M (Art No 10-9529-01: for 6 x 4 determinations) • ImmunoCAP Specific IgE Control H (Art No 10-9530-01: for 6 x 4 determinations) • ImmunoCAP Specific IgE Negative Control (Art No 10-9445-01: for 6 x 4 determinations)Reagents for Phadia 1000• ImmunoCAP Specific IgE Conjugate 100 (Art No 10-9316-01: for 6 x 100 determinations) • ImmunoCAP Specific IgE Conjugate 400 (Art No 10-9310-01: for 6 x 400 determinations) • ImmunoCAP Specific IgE Calibrator Strip 0-100 (Art No 10-9459-01: for 5 calibration curves) • ImmunoCAP Specific IgE Curve Control Strip (CC-1 and CC-2) (Art No 10-9312-01: 5 x 3 sets of curve control) • ImmunoCAP Specific IgE Anti-IgE (a_IgE) (Art No 14-4417-01: carriers of 16 ImmunoCAP) • ImmunoCAP Allergen and Allergen Mix (See Product catalogue: carriers of 16 or 10 ImmunoCAP) • ImmunoCAP Total IgE Low Range (low) (Art No 14-4497-35: for 48 determinations) • ImmunoCAP Phadiatop (phad) (Art No 14-4405-35: for 48 determinations) • ImmunoCAP Phadiatop Infant (phinf) (Art No 14-4510-35: for 48 determinations) • Development Solution (Art No 10-9439-01: for 6 x 1200 determinations; Art No 10-9314-01: for 6 x 2000 determinations) • Stop Solution (Art No 34-2271-51: for 1200 determinations) • Washing Solution (Art No 10-9202-01: 2 x 5 l) Washing Solution Additive, 2 x 86 ml• • Washing Solution Concentrate, 2 x 400 ml • ImmunoCAP Specific IgE Control L (Art No 10-9528-01: for 6 x 4 determinations) • ImmunoCAP Specific IgE Control M (Art No 10-9529-01: for 6 x 4 determinations) • ImmunoCAP Specific IgE Control H (Art No 10-9530-01: for 6 x 4 determinations) • ImmunoCAP Specific IgE Negative Control (Art No 10-9445-01: for 6 x 4 determinations)Reagents for Phadia 2500 and Phadia 5000• ImmunoCAP Specific IgE Conjugate 400 (Art No 10-9310-01: for 6 x 400 determinations) • ImmunoCAP Specific IgE Calibrator Strip 0-100 (Art No 10-9459-01: for 5 calibration curves) • ImmunoCAP Specific IgE Curve Control Strip (CC-1 and CC-2) (Art No 10-9312-01: 5 x 3 sets of curve control) • ImmunoCAP Specific IgE Anti-IgE (a_IgE) (Art No 14-4417-01: carriers of 16 ImmunoCAP) • ImmunoCAP Allergen and Allergen Mix (See Product catalogue: carriers of 16 or 10 ImmunoCAP) • ImmunoCAP Total IgE Low Range (low) (Art No 14-4497-35: for 48 determinations) • ImmunoCAP Phadiatop (phad) (Art No 14-4405-35: for 48 determinations) • ImmunoCAP Phadiatop Infant (phinf) (Art No 14-4510-35: for 48 determinations) • Development Solution (Art No 10-9314-01: for 6 x 2000 determinations) • Stop Solution (Art No 34-2337-11: for 4600 determinations) • Washing Solution Additive (Art No 10-9518-01: 4 x 850 ml) • Washing Solution Concentrate (Art No 34-2337-21: 1 x 2800 ml) • ImmunoCAP Specific IgE Control L (Art No 10-9528-01: for 6 x 4 determinations) • ImmunoCAP Specific IgE Control M (Art No 10-9529-01: for 6 x 4 determinations) • ImmunoCAP Specific IgE Control H (Art No 10-9530-01: for 6 x 4 determinations) • ImmunoCAP Specific IgE Negative Control (Art No 10-9445-01: for 6 x 4 determinations)Details of reagents ImmunoCAP Specific IgE Conjugate 100/Conjugate 400Ready for use. Store at 2 – 8 °C until expiration date. Do not freeze!ß-Galactosidase-anti-IgE Approximately 1 μg/ml (mouse monoclonal antibodies) Sodium azide 0.06%ImmunoCAP Specific IgE Calibrator Strip 0-100Ready for use. Store at 2 – 8 °C until expiration date. (human IgE in buffer) Conc. 0; 0.35; 0.7; 3.5; 17.5 and 100 kU/l Preservative* <0.003%ImmunoCAP Specific IgE Curve Control StripReady for use. Store at 2 – 8 °C until expiration date. (human IgE in buffer) Preservative* <0.003%ImmunoCAP Specific IgE Anti-IgEReady for use. Store at 2 – 8 °C until expiration date. (mouse monoclonal antibodies) Preservative* <0.0015%ImmunoCAP Allergen or Allergen MixReady for use. Store at 2 – 8 °C until expiration date. Preservative* <0.0015%ImmunoCAP Total IgE Low RangeFor information, see separate Directions for Use for ImmunoCAP Total IgE Low Range.ImmunoCAP PhadiatopFor information, see separate Directions for Use for ImmunoCAP Phadiatop.ImmunoCAP Phadiatop InfantFor information, see separate Directions for Use for ImmunoCAP Phadiatop Infant.Development SolutionReady for use. Store at 2 – 8 °C until expiration date. Do not freeze!4-Methylumbelliferyl-ß-D-galactoside 0.01% Preservative* <0.0010% 52-5291-EN/12 Page 1 (4)Published 2025-09-30 ### Page 2 Stop SolutionReady for use. Store at 2 – 32 °C until expiration date. Sodium carbonate 4%Washing SolutionFor information, see separate Directions for Use for Washing Solution.ImmunoCAP Specific IgE Control LReady for use. Store at 2 – 8 °C until expiration date. Sodium azide 0.05% ImmunoCAP Specific IgE Control L is prepared from selected pooled human samples and contains IgE antibodies to the allergen e1.ImmunoCAP Specific IgE Control MReady for use. Store at 2 – 8 °C until expiration date. Sodium azide 0.05% ImmunoCAP Specific IgE Control M is prepared from selected pooled human samples and contains IgE antibodies to the allergen t3.ImmunoCAP Specific IgE Control HReady for use. Store at 2 – 8 °C until expiration date. Sodium azide 0.05% ImmunoCAP Specific IgE Control H is prepared from selected pooled human samples and contains IgE antibodies to the allergen d1.ImmunoCAP Specific IgE Negative ControlReady for use. Store at 2 – 8 °C until expiration date. Sodium azide 0.05% ImmunoCAP Specific IgE Negative Control is prepared from selected pooled human samples.ImmunoCAP IgE/ECP/Tryptase Sample DiluentReady for use. Store at 2 – 8 °C until expiration date. (buffer solution with Bovine Serum Albumin) Preservative* <0.003%*Preservative: Reaction mass of CMIT/MIT (3:1), (CAS No: 55965-84-9).Additional materialAdditional products available from Phadia AB: • ImmunoCAP IgE/ECP/Tryptase Sample Diluent (Art No 10-9360-01/10-9541-01)PrecautionsWarning• For in vitro diagnostic use. Not for internal or external use in humans or animals. • Some reagents are manufactured from human blood components. The source materials have been tested by immunoassay for hepatitis B surface antigen, for antibodies to HIV1, HIV2 and hepatitis C virus and found to be negative. Nevertheless, all recommended precautions for the handling of blood derivatives should be observed. Please refer to Human Health Service (HHS) Publication No. (CDC) 93-8395 or other local/national guidelines on laboratory safety procedures. • Reagents containing ≥0.00015% reaction mass of CMIT/MIT (3:1) (CAS No: 55965-84-9) may produce an allergic reaction (EUH 208). For more information see Safety Data Sheet. • Reagents containing ≥0.0015% reaction mass of CMIT/MIT (3:1) (CAS No: 55965-84-9) may cause an allergic skin reaction (H317). Wear protective gloves/protective clothing/eye protection (P280). Gloves: Nitrile rubber EN374. For more information see Safety Data Sheet. • Reagents containing ≥0.0025% reaction mass of CMIT/MIT (3:1) (CAS No: 55965-84-9) are classified as harmful to aquatic life with long lasting effects (H412). Avoid release to the environment (P273). Dispose of contents/container in accordance with local/national regulations (P501). For more information see Safety Data Sheet.DisposalFollow local disposal regulations based on your location along with recommendations and content in the Safety Data Sheet to determine the safe disposal of the products.INSTRUMENTSPhadia 200 processes all steps of the assay and prints results automatically after the assay is completed(a). Phadia 250, Phadia 1000, Phadia 2500 and Phadia 5000 are continuous random access systems that perform all steps of the assay(a).On board stabilityInformation on reagent storage for the intended locations on the Phadia instruments is shown below.Phadia 2500, Phadia 5000 Phadia 1000Phadia 250Phadia 200N/A28 days.Calibrator/ Curve Control7 days.4 days.ConjugateRecap bottles every night. 5 days.14 days.5 days.Development Solution Recap bottles every night. 31 days.14 days.14 days.Stop SolutionRecap bottles every night. 7 days.Washing Solution(prepared solution)Discard every seventh day and perform weekly maintenance according to respective instrument user manual. 30 days.N/AWashing Solution Concentrate30 days.N/AWashing Solution AdditiveN/A7 days.Sample Diluent Recap bottles every night. Until expiration date.No provision for on board storage.ImmunoCAP Carrier SPECIMEN COLLECTION AND PREPARATION Specimen collectionSerum and plasma (EDTA or heparin) samples from venous or capillary blood can be used. Collect and treat blood samples using standard procedures. Store at 2 °C to 8 °C up to one week, or else at –20 °C. Storage at room temperature is not recommended. However, for shipping purposes, specimens may be kept at room temperature (not exceeding 32 °C) for up to one week(b). Avoid repeated freezing and thawing (13). For further reading on interfering substances and IgE stability in specimens, see references (14-15).Note: It is the responsibility of the individual laboratory to use all available references and/or its own studies to determine specific stability criteria for its laboratory. In general, laboratories should perform validation studies before implementing a change in specimen acceptance criteria (16).Note: Blood samples for testing of specific IgE to a drug or venom should be collected as soon as possible after a known exposure or clinical reaction (17, 18), preferably not later than 6 months after the event (18-20). If the test result is negative and an IgE-mediated reaction is still suspected, it has been recommended to confirm the result by drawing a new sample and repeating the analysis (21-23).Preparation of samplesSample dilution is usually not required. For determination of values higher than 100 kUA/l IgE, dilute the samples with ImmunoCAP IgE/ECP/Tryptase Sample Diluent.Handling of control specimenIt is recommended to remove and recap the control vials from the instrument as soon as the pipetting of the samples is finished and the sample incubation is started. It is also recommended to gently stir the vial before use. ImmunoCAP Specific IgE Controls should be treated in the same way as a patient sample in the procedure. ImmunoCAP Specific IgE Controls are ready for use and must not be further diluted.PROCEDURES Procedural steps(a)For procedural steps, see Notes a.Parameters of the procedurePatient samples are run in single determinations. Volumes per determination: 40 μlSample 50 μlConjugate 50 μlDevelopment Solution 600 μlStop Solution Incubations are performed at 37 °C by Phadia instruments. Phadia 200: Total time for one assay is up to 4 hours. Phadia 250, Phadia 1000, Phadia 2500 and Phadia 5000: The process time is 1 hour and 45 minutes from entering the first sample until the result is available.Calibration(a)The calibrators, provided in ImmunoCAP Specific IgE Calibrator Strip, are run in duplicates to obtain a full calibration curve. The calibration curve will be stored for use in subsequent assay runs. The two curve controls CC1 and CC2, each in single determinations, are used to approve the validity of the previously stored calibration curve. The software for Phadia instruments have built-in acceptance limits for the calibration curve and the curve controls.Calibrator range: 0-100 kU/l.Reference material: The IgE calibrators are traceable (via an unbroken chain of calibrations) to the 3rd International Standard 11/234, of Human Serum Immunoglobulin E from World Health Organization (WHO) (24).Accuracy(b): Measured values of ImmunoCAP Specific IgE are, with 95% probability, within ±8% of the WHO IgE reference value.Trueness(b): The bias of ImmunoCAP Specific IgE Calibrators is, with 95% probability, within ±2% compared to the WHO IgE reference value.QUALITY CONTROL Record keeping for each assayIt is good laboratory practice to record the lot numbers of the components used, the dates when they were first opened and the remaining volumes.Control specimenGood laboratory practice requires that quality control specimen should be included in every run. Any material used should be assayed repeatedly to establish mean values and acceptable ranges. Controls available from Phadia AB for day to day quality control: • ImmunoCAP Specific IgE Control L (Art No 10-9528-01) • ImmunoCAP Specific IgE Control M (Art No 10-9529-01) • ImmunoCAP Specific IgE Control H (Art No 10-9530-01) • ImmunoCAP Specific IgE Negative Control (Art No 10-9445-01)Intended use for ImmunoCAP Specific IgE ControlsImmunoCAP Specific IgE Controls are used for monitoring ImmunoCAP Specific IgE measurements performance in Phadia instruments.Expected values for positive ImmunoCAP Specific IgE ControlsAs with all immunoassays the results are affected by the testing procedures and equipment used by different laboratories. It is therefore recommended that each laboratory establishes its own target value for each actual lot of control together with criteria of acceptance (recommended range ±30%). 52-5291-EN/12 Page 2 (4)Published 2025-09-30 ### Page 3 This established target value is expected to fall within the range for the actual lot. The range is stated on the label of each ImmunoCAP Specific IgE Control vial. The range for each specific lot is the mean concentration ±2 standard deviations (SD), where the mean value has been determined from at least 8 consecutive assays, each in a minimum of 6 replicates using ImmunoCAP Specific IgE. The standard deviation is based on the expected long-term variation for the specific IgE assay.Expected values for ImmunoCAP Specific IgE Negative ControlThe negative control will give results representative for non-atopic blood donors using any of the following ImmunoCAP Allergens: • ImmunoCAP Allergen e1, Cat dander (Art No 14-4109-01) • ImmunoCAP Allergen t3, Common silver birch (Art No 14-4102-01) • ImmunoCAP Allergen d1, House dust mite (Art No 14-4107-01) Expected values are below 0.1 kUA/l.Proficiency testingAn external quality assessment program (proficiency testing) is available from Phadia AB for quality assurance purposes (Quality Club): • Quality Club Specific IgE (Art No 10-9298-01)RESULTSFor Phadia instruments using Phadia Prime or Phadia Information Data Manager software, all calculations are automatically performed(a).Interpretation of results for ImmunoCAP AllergensQuantitative evaluation of specific IgE antibody concentration (kUA/l)For ImmunoCAP Allergens, results are expressed as concentration in kUA/l, where A represents allergen-specific IgE antibodies. Values above the limit of quantitation represent a progressive increase in the concentration of allergen-specific antibodies. For interpretation of results for the following products, see their respective DFUs: • ImmunoCAP Phadiatop (Art No 14-4405-35) • ImmunoCAP Phadiatop Infant (Art No 14-4510-35) • ImmunoCAP Total IgE Low Range (Art No 14-4497-35)Interpretation of results for ImmunoCAP Allergen MixesQualitative evaluation of specific IgE antibodiesThe results from ImmunoCAP Allergen Mixes are expressed as positive or negative, with 0.35 kUA/l as analytical cut-off. A positive result indicates specific IgE antibodies to one or more of the allergens in the ImmunoCAP Allergen Mix. A negative result indicates undetectable levels of allergen-specific IgE antibodies. Deviations from results obtained with ImmunoCAP Allergen tests may occur and a negative result for an ImmunoCAP Allergen Mix does not exclude allergy sensitization. Results obtained with an ImmunoCAP Allergen Mix do not correspond to the cumulative results obtained with ImmunoCAP Allergens representing the allergen sources combined in the ImmunoCAP Allergen Mix.LIMITATIONS OF THE PROCEDUREA definitive clinical diagnosis should only be made by the physician after all clinical and laboratory findings have been evaluated. It should not be based on the results of any single diagnostic method. • Very low levels of allergen-specific IgE antibodies should be evaluated with caution when: a. total IgE values are above 1,000 kU/l b. total IgE values are above 500 kU/l when testing for specific IgE antibodies to beta-lactams and chlorhexidine (ImmunoCAP Allergen c1, c2, c5, c6, c7 & c8) • In food allergy, circulating IgE antibodies may remain undetectable despite a convincing clinical history. The antibodies may be directed towards allergens that are revealed or altered during industrial processing, cooking or digestion and therefore do not exist in the original food for which the patient is tested. • Results below limit of quantitation obtained for a drug- or venom specific IgE determination indicates the absence of specific IgE antibodies to that drug or venom. Such results do not preclude existence of current or future clinical hypersensitivity to drugs or venoms. • Samples with results below limit of quantitation obtained with ImmunoCAP Allergen Components are recommended to be tested with the corresponding extract-based ImmunoCAP Allergen, if not already performed. Such complementary extract-based testing will cover additional allergen components present in the allergen source material to which the patient may be sensitized. • A result below limit of quantitation obtained with an extract-based ImmunoCAP Allergen does not exclude the possibility of obtaining measurable concentrations of specific IgE when testing with ImmunoCAP Allergen Components from the same allergen source. This is due to the fact that some components may be present in very low amounts in the natural extract.EXPECTED VALUES(b)Good laboratory practice recommends that each laboratory establishes its own expected range of values (25). There is no defined clinical cut-off value for allergen-specific IgE. In healthy non-sensitized individuals, the expected value for allergen-specific IgE antibodies is below 0.1 kUA/l (1). Values above 0.1 kUA/l represent a risk of symptoms upon allergen exposure (13, 26). Studies have shown an association between increasing levels of allergen-specific IgE antibodies and higher probability of allergic symptoms upon exposure to the corresponding allergen (27-30). The ability to measure allergen-specific IgE antibodies down to 0.1 kUA/l has been shown to be clinically relevant (31-34), although levels between 0.1 and 0.35 kUA/l are often not associated with clinically evident disease and should be cautiously interpreted (13). The presence of IgE antibodies of defined specificity should be considered as a risk factor for allergic disease and viewed in the context of information obtained from the clinical evaluation of the patient and other diagnostic procedures in making a diagnosis of allergic disease (13). In clinical practice, 0.35 kUA/l has commonly been used as a cut off and a large number of studies have been performed in which the clinical performance of ImmunoCAP Specific IgE tests in allergy diagnosis has been evaluated. A meta-analysis of six independent studies including 894 patients tested for IgE antibodies to a range of different allergens using ImmunoCAP Specific IgE showed an overall clinical sensitivity of 89% (84-95% for the different studies) and specificity of 91% (85-94% for the different studies) (35).PERFORMANCE CHARACTERISTICS(b)Instrument comparisonIn an instrument comparison study with 196 patient samples and 50 ImmunoCAP Allergens, the following average differences between each Phadia instrument system and the overall mean were estimated:Average difference to mean (%)Phadia instrument-3.3Phadia 200 -0.6Phadia 250 -0.6Phadia 1000 4.7Phadia 2500/5000Analytical specificityNo interference of rheumatoid factor (<400 IU/ml), hemoglobin (<5 g/l), triglycerides (chyle <1,490 FTU) or bilirubin (bilirubin coupled <21 mg/dl, bilirubin free <19 mg/dl) has been observed in studies, when tested at recommended test concentrations (36). The ImmunoCAP assay system has been shown to be sufficiently robust to provide accurate and reproducible specific IgE results in specimens containing therapeutic concentrations of omalizumab (37). Cross-reactivity with other human immunoglobulins is non-detectable at physiological concentrations of IgA, IgD, IgM and IgG.ImmunoCAP AllergensPrecisionThe following pooled coefficients of variation have been obtained in a study with Phadia 250 when testing representative allergens from 8 allergen groups, in total 4-8 samples/level. Each sample has been assayed in 4 replicates on 7 different occasions on each of 3 different instruments, using the same lot of reagents and stored calibration curves. The values are representative for all Phadia instruments.Coefficients of variation (%)Sample level (kUA/l) Between assayWithin assay56LoQ – 0.35 640.35 – 1.5 451.5 – 50 7850 – 100Limit of quantitationThe overall limit of quantitation (38) for allergen-specific IgE antibodies is 0.1 kUA/l.LinearityThe overall dilution linearity (13) in the concentration range of 0.1 – 100 kUA/l has been demonstrated in studies performed on representative allergens from 11 allergen groups and 3 samples/allergen. The average concentration range covered for the allergens was 0.13 – 64 kUA/l. The relationship between observed and expected concentration was evaluated with linear regression analysis (estimated on log10 concentration). The estimated slopes were within 0.89 – 1.07 and the intercepts were within -0.14 – 0.24.ImmunoCAP Allergen MixesPrecisionFor ImmunoCAP Allergen Mixes, the precision was evaluated near the cut-off level. Nine representative ImmunoCAP Allergen Mixes were tested using 5 samples, all giving results near 0.35 kUA/l. Each sample was assayed in 5 replicates on 3 different occasions on two reagents lots using one Phadia 1000 instrument. The precision results were expressed in terms of C5 (the concentration below cut-off where repeated testing of a sample is expected to give positive results in 5% of the cases) and C95 (the concentration above cut-off where repeated testing of a sample is expected to give positive results in 95% of the cases). In this study, C5 was estimated to 0.31 kUA/l and C95 was estimated to 0.39 kUA/l (39).Limit of detectionThe overall limit of detection (38) for allergen-specific IgE antibodies is below calibrator value 0.35 kU/l.WARRANTYThe performance data presented here was obtained using the procedure indicated. Any change or modification in the procedure not recommended by Phadia AB may affect the results, in which event Phadia AB disclaims all warranties expressed, implied or statutory, including the implied warranty of merchantability and fitness for use. Phadia AB and its authorized distributors, in such event, shall not be liable for damages indirect or consequential.IMPORTANT NOTICEFor healthcare professionals in the European Union and in countries with identical regulatory regime (Regulation 2017/746/EU on In vitro Diagnostic Medical Devices); if, during the use of this device or as a result of its use, a serious incident has occurred, please report it to the manufacturer and/or its authorized representative and to your national authority.SYMBOLSIn vitro diagnostic medical deviceUse-by dateTemperature limitBatch codeConsult instructions for useDate of manufactureBiological risksCatalogue numberFor prescription use only – only applicable under US legislationRx onlyCautionWarningManufacturerContains sufficient for testsΣFull symbol glossary is available at: https://symbols_glossary.phadia.com.REFERENCES1. Johansson SG, Bieber T, Dahl R, Friedmann PS, Lanier BQ, Lockey RF, et al. Revised nomenclature for allergy for global use: Report of the Nomenclature Review Committee of the World Allergy Organization, October 2003. J Allergy Clin Immunol 2004;113(5):832-6. 2. Ishizaka K, Ishizaka T, Hornbrook MM. Physico-chemical properties of human reaginic antibody. IV. Presence of a unique immunoglobulin as a carrier of reaginic activity. J Immunol 1966;97(1):75-85. 3. Wide L, Bennich H, Johansson SG. Diagnosis of allergy by an in-vitro test for allergen antibodies. Lancet 1967;2(7526):1105-7. 4. Yunginger JW, Ahlstedt S, Eggleston PA, Homburger HA, Nelson HS, Ownby DR, et al. Quantitative IgE antibody assays in allergic diseases. J Allergy Clin Immunol 2000;105(6 Pt 1):1077-84. 5. Ahlstedt S. Understanding the usefulness of specific IgE blood tests in allergy. Clin Exp Allergy 2002;32(1):11-6. 6. Wahn U. What drives the allergic march? Allergy 2000;55(7):591-9. 52-5291-EN/12 Page 3 (4)Published 2025-09-30 ### Page 4 7. Sicherer SH, Sampson HA. Cow's milk protein-specific IgE concentrations in two age groups of milk-allergic children and in children achieving clinical tolerance. Clin Exp Allergy 1999;29(4):507-12. 8. Crespo JF, Pascual C, Ferrer A, Burks AW, Diaz Pena JM, Martin Esteban M. Egg white-specific IgE level as a tolerance marker in the follow up of egg allergy. Allergy Proc 1994;15(2):73-6. 9. Boyano-Martinez T, Garcia-Ara C, Diaz-Pena JM, Martin-Esteban M. Prediction of tolerance on the basis of quantification of egg white-specific IgE antibodies in children with egg allergy. J Allergy Clin Immunol 2002;110(2):304-9. 10. Shek LP, Soderstrom L, Ahlstedt S, Beyer K, Sampson HA. Determination of food specific IgE levels over time can predict the development of tolerance in cow's milk and hen's egg allergy. J Allergy Clin Immunol 2004;114(2):387-91. 11. Shirai T, Matsui T, Suzuki K, Chida K. Effect of pet removal on pet allergic asthma. Chest. 2005;127(5):1565-71. 12. Hamilton RG, Williams PB. Human IgE antibody serology: a primer for the practicing North American allergist/immunologist. J Allergy Clin Immunol. 2010;126(1):33-8. 13. CLSI. Analytical Performance Characteristics, Quality Assurance, and Clinical Utility of Immunological Assays for Human Immunoglobulin E Antibodies of Defined Allergen Specificities; Approved Guideline - Third Edition. CLSI document I/LA20. Wayne, PA: Clinical and Laboratory Standards Institute; 2016. 14. Tate J, Ward G: Interferences in immunoassay. Clin Biochem Rev. 2004:25:105-120. 15. World Health Organization. Diagnostic Imaging and Laboratory Technology. (2002). Use of anticoagulants in diagnostic laboratory investigations. World Health Organization. WHO/DIL/LAB/99.1 Rev.2. 16. CLSI. Procedures for the Handling and Processing of Blood Specimens for Common Laboratory Tests; Approved Guideline - Fourth Edition. CLSI document GP44-A4, Wayne, PA: Clinical and Laboratory Standards Institute; 2010. 17. Guttormsen AB, Johansson SGO, Öman H, Wilhelmsen V, Nopp A. No consumption of IgE antibody in serum during allergic drug anaphylaxis. Allergy 2007; 62: 1326-1330. 18. Garvey LH, Kroigaard M, Poulsen LK, Skov PS, Mosbech H, Venemalm L, et al. IgE-mediated allergy to chlorhexidine. J Allergy Clin Immunol. 2007; 120(2):409-15. 19. Mosbech H, Christensen J, Dirksen A, Soborg M. Insect allergy. Predictive value of diagnostic tests: a three-year follow-up study. Clin Allergy. 1986;16(5):433-40. 20. Hjortlund J, Mortz CG, Stage TB, Skov PS, Dahl R, Bindslev-Jensen C. Positive serum specific IgE has a short half-life in patients with penicillin allergy and reversal does not always indicate tolerance. Clin Transl Allergy. 2014;4:34. 21. Rieger-Ziegler V, Rieger E, Kränke B, Aberer W. Hymenoptera venom allergy: time course of specific IgE concentrations during the first weeks after a sting. Int Arch Allergy Immunol. 1999;120(2):166-8. 22. Goldberg A, Confino-Cohen R. Timing of venom skin tests and IgE determinations after insect sting anaphylaxis. J Allergy Clin Immunol. 1997;100(2):182-4. 23. Kraft D, Wide L. Clinical patterns and results of radioallergosorbent test (RAST) and skin tests in penicillin allergy. Br J Dermatol. 1976;94(6):593-601. 24. Thorpe SJ, Heath A, Fox B, Patel D, Egner W. The 3rd International Standard for serum IgE: international collaborative study to evaluate a candidate preparation. Clin Chem Lab Med. 2014; 52(9): 1283-9. 25. CLSI. Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; Approved Guideline - Third Edition. CLSI document EP28-A3C. Wayne, PA, USA: Clinical and Laboratory Standards Institute; 2008. 26. Hamilton RG. Clinical laboratories worldwide need to report IgE antibody results on clinical specimens as analytical results and not use differential positive thresholds. J Allergy Clin Immunol. 2015;135:811-2. 27. Celik-Bilgili S, Mehl A, Verstege A, Staden U, Nocon M, Beyer K, et al. The predictive value of specific immunoglobulin E levels in serum for the outcome of oral food challenges. Clin Exp Allergy. 2005;35(3):268-73. 28. Simpson A, Soderstrom L, Ahlstedt S, Murray CS, Woodcock A, Custovic A. IgE antibody quantification and the probability of wheeze in preschool children. J Allergy Clin Immunol. 2005;116(4):744-9. 29. Maloney JM, Rudengren M, Ahlstedt S, Bock SA, Sampson HA. The use of serum-specific IgE measurements for the diagnosis of peanut, tree nut, and seed allergy. J Allergy Clin Immunol. 2008;122(1):145-51. 30. Beyer K, Grabenhenrich L, Hartl M, Beder A, Kalb B, Ziegert M, et al. Predictive values of component-specific IgE for the outcome of peanut and hazelnut food challenges in children. Allergy. 2015;70(1):90-8. 31. Ebo DG, Venemalm L, Bridts CH, Degerbeck F, Hagberg H, De Clerck LS, et al. Immunoglobulin E antibodies to rocuronium: a new diagnostic tool. Anesthesiology. 2007;107(2):253-9. 32. Guerti K, Bridts CH, Stevens WJ, Ebo DG. Wasp venom-specific IgE: towards a new decision threshold? J Investig Allergol Clin Immunol. 2008;18(4):321-3. 33. Soderstrom L, Lilja G, Borres MP, Nilsson C. An explorative study of low levels of allergen-specific IgE and clinical allergy symptoms during early childhood. Allergy. 2011;66(8):1058-64. 34. Linden CC, Misiak RT, Wegienka G, Havstad S, Ownby DR, Johnson CC, et al. Analysis of allergen specific IgE cut points to cat and dog in the Childhood Allergy Study. Ann Allergy Asthma Immunol. 2011;106(2):153-8 e2. 35. Paganelli R, Ansotegui IJ, Sastre J, Lange CE, Roovers MH, de Groot H, et al. Specific IgE antibodies in the diagnosis of atopic disease. Clinical evaluation of a new in vitro test system, UniCAP, in six European allergy clinics. Allergy 1998;53(8):763-8. 36. CLSI. Interference Testing in Clinical Chemistry; Approved Guideline - Second Edition. CLSI document EP07-A2. Wayne, PA: Clinical and Laboratory Standards Institute; 2005. 37. Hamilton RG. Accuracy of US Food and Drug Administration-cleared IgE antibody assays in the presence of anti-IgE (omalizumab). J Allergy Clin Immunol. 2006 Apr;117(4):759-66. 38. NCCLS EP17-A. Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Guideline – First Edition, National Committee for Clinical Laboratory Standards, 2004. 39. CLSI. User Protocol for Evaluation of Qualitative Test Performance; Approved Guideline - Second Edition. CLSI document EP12-A2. Wayne; PA: Clinical and Laboratory Standards Institute; 2008.Notes(a)For more information, see Phadia 200, Phadia 250, Phadia 1000 and/or Phadia 2500/Phadia 5000 user manual.(b)Studies performed at Phadia AB, Uppsala, Sweden.Patents/TrademarksThe following designations are trademarks belonging to Phadia AB: ImmunoCAP, Phadia, Phadiatop, Quality Club.Trademark change: Phadia AB has changed the trademarks of the instrument platforms from "UniCAP™" and "ImmunoCAP™" to "Phadia™". The new name has been applied to the instruments and related items, e.g. software and user manuals. The trademark "ImmunoCAP™" has been removed from the System Reagents. This is a trademark change only; the change has no impact on performance or safety.Addresseshttp://www.thermofisher.com/phadia-locations Phadia AB,Rapsgatan 7P, P. O. Box 6460, 751 37 Uppsala, Sweden Tel: +46 18 16 50 00 Fax: +46 18 14 03 58 0459Issued October 2011. Revised May 2025. © Phadia AB, Uppsala, Sweden.Revision history Version 12Removal of Phadia 100 instrument and Phadia 100-associated reagents throughout the document. Rephrasing of texts in: Calibration. Updated calculations for: Instrument comparison.Addresses: Removal of addresses to country offices. Addition of website link to: Phadia Commercial Organization and Distributor Locations. 52-5291-EN/12 Page 4 (4)Trace back: Workspace: DFU ImmunoCAP Edition a422, Frozen, Publication: 52-5291-EN_ImmunoCAP_Specific_IgEPublished 2025-09-30