Please only refer to the current product lot insert enclosed with the kits package for execution

and reporting

0123 130203004M: 100 tests

130603004M: 050 tests

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MAGLUMI FT4 (CLIA)
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INTENDED USE
The kit is an in vitro chemiluminescence immunoassay for the quantitative determination of Free Thyroxine (FT4) in human serum using the
MAGLUMI series Fully-auto chemiluminescence immunoassay analyzer (including Maglumi 600, Maglumi 800, Maglumi 1000, Maglumi 1000
Plus, Maglumi 2000, Maglumi 2000 Plus, Maglumi 4000 and Maglumi 4000 Plus).

SUMMARY AND EXPLANATION OF THE TEST

Thyroxine (T4), a thyroid hormone with the highest concentration in the body, is generated in the thyroid gland by coupling two di-iodinated tyrosine
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molecules contained within the thyroglobulin molecule. Together with other thyroid hormones such as triiodothyronine (T3), it exerts a great
influence on the activities of the body, including the basal metabolic rate, protein synthesis, regulation of bone growth, neuronal maturation and
sensitivity to other hormones1.
T4 circulates in the bloodstream as an equilibrium mixture of bound and unbound form, where approximately 99.97% is bound to plasma proteins:
thyroxine binding globulin (TBG) 60-70%, transthyretin or thyroxine binding prealbumin (TTR/TBPA) 15-30%, and albumin 10%, however the
remaining 0.03% unbound T4 named FT4 is substantially physiologically active to serve as metabolic stimulation and control feedback system of
hypothalamic-pituitary-thyroid axis2,3. The equilibrium is always maintained to guarantee the constant level of free T4 in spite of alterations occur in
either the concentration or affinity of the serum binding proteins, therefore FT4 is a more reliable indicator of an individual’s thyroid status than total
T4 due to its less sensitivity to changes in serum binding proteins4,5.
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Free T4 is elevated in hyperthyroidism and depressed in hypothyroidism, and more additional thyroid tests such as free T3 and TSH may be very
indispensible in borderline cases of suspected thyroid malfunction.

PRINCIPLE OF THE TEST

The FT4 assay is a competitive chemiluminescence immunoassay.
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The sample(or calibrator/control, if applicable), ABEI-labeled anti-T4 monoclonal antibody, buffer and T4 antigen-coated magnetic microbeads
are mixed thoroughly and incubated at 37°C. Free T4 present in the serum sample (or calibrator/control, if applicable) competes with T4 antigen
immobilized on the magnetic microbeads for a limited number of binding sites on the ABEI-labeled anti-T4 antibody, forming immuno-complexes.
After precipitation in a magnetic field, decant the supernatant, and then perform a wash cycle. Subsequently, the Starter 1+2 are added to initiate a
chemiluminescent reaction. The light signal is measured by a photomultiplier within 3 seconds as relative light units (RLUs), which is inversely
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proportional to the concentration of free T4 present in the sample (or calibrator/control, if applicable).

KIT COMPONENTS
Material Provided
100 tests 50 tests
Component Contents
(REF:130203004M) (REF:130603004M)
Magnetic Magnetic microbeads coated with purified T4 antigen,
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2.5 mL 2.0 mL
Microbeads containing BSA, NaN3 (<0.1%).
Calibrator Low Containing BSA and T4 antigen, NaN3 (<0.1%). 2.5 mL 2.0 mL
Calibrator High Containing BSA and T4 antigen, NaN3 (<0.1%). 2.5 mL 2.0 mL
Buffer Containing BSA, NaN3 (<0.1%). 6.5 mL 4.0 mL
Anti-T4 monoclonal antibody labeled with ABEI,
ABEI Label 6.5 mL 4.0 mL
containing BSA, NaN3 (<0.1%).
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Internal Quality
Containing BSA and T4 antigen, NaN3(<0.1%). 2.0 mL 2.0 mL
Control
All reagents are provided ready-to-use.

Accessories Required But Not Provided

MAGLUMI Series:
Reaction Module REF: 630003
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Starter 1+2 REF: 130299004M

Wash Concentrate REF: 130299005M
Light Check REF: 130299006M
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Please order accessories from Shenzhen New Industries Biomedical Engineering Co., Ltd. (SNIBE) or our authorized representatives.

CALIBRATION
Traceability: This method has been standardized against USP (United States Pharmacopeia) Standards.
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Test of assay specific calibrators allows the RLU values to adjust the assigned master curve. Results are determined via a calibration curve which is
instrument-specifically generated by 2-point calibrationand a master curve (10 calibrations) provided via the reagent Radio Frequency Identification
(RFID) CHIP.
Recalibration is recommended if any of the following conditions occurs:
 After each exchange of lots (Reagent or Starter 1+2).
 Every week and/or each time a new reagent kit is used (recommended).
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 After instrument service is required.

 If controls lie outside the expected range.
 Whenever room temperature changes exceed 5° C (recommended).

QUALITY CONTROL
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Follow government regulations or accreditation requirements for quality control frequency.

Internal quality control is only applicable with MAGLUMI system. For instructions for use and target value refer to FT4 (CLIA) Quality Control
Information. User needs to judge results with their own standards and knowledge.
For detailed information about entering quality control values, refer to the operating instructions of MAGLUMI series Fully-auto chemiluminescence
immunoassay analyzer.
To monitor system performance and chart trends, commercially available quality control materials are required. Treat all quality control samples the
same as patient samples. A satisfactory level of performance is achieved when analyte values obtained are within the acceptable Control Range for
the system or within your range, as determined by an appropriate internal laboratory quality control scheme. If the quality control results do not fall
within the Expected Values or within the laboratory’s established values, do not report results. Take the following actions:
 Verify that the materials are not expired.
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 Verify that required maintenance was performed.
 Verify that the assay was performed according to the instructions for use.
 Rerun the assay with fresh quality control samples.
 If necessary, contact your local technical supporters or distributors for assistance.

SPECIMEN COLLECTION AND PREPARATION

 Use standard sampling tubes or tubes containing separating gel. Collect blood aseptically followed the universal precautions for venipuncture.
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 To ensure consistency in results, specimens must be transferred to a centrifuge tube and centrifuged at ≥10,000 RCF (Relative Centrifugal
Force) for 15 minutes.
 Ensure that complete clot formation in serum specimens has taken place prior to centrifugation. Some specimens, especially those from patients
receiving anticoagulant or thrombolytic therapy, may exhibit increased clotting time.
 If the specimen is centrifuged before a complete clotting, the presence of fibrin may cause erroneous results. Samples must be free of fibrin and
other particulate matter.
 Do not use hemolyzed or grossly lipemic specimens as well as specimens containing particulate matter or exhibiting obvious microbial
contamination. Inspect all specimens for bubbles, and remove bubbles before analysis for optimal results.
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 Avoid repeating freeze-thaw cycles. The serum sample can be only frozen and thawed once. Specimens must be mixed thoroughly after thawing.
 Centrifuged specimens with a lipid layer on the top must be transferred to a sample cup or a secondary tube. Care should be taken to transfer
only the clarified specimen without the lipemic material.
 All samples (patient specimens and controls) should be tested within 3 hours when placed on board the MAGLUMI System. Refer to the SNIBE
service for more detailed discussion of onboard sample storage constraints.
 If testing will be delayed for more than 8 hours, remove serum from the serum separator, red blood cells or clot. Specimens removed from the
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separator, cells or clot may be stored up to 24 hours at 2-8°C. Freeze samples at or below -20°C if the sample is not assayed within 24 hours.
 Before shipping specimens, it is recommended that specimens be removed from the serum separator, red blood cells or clot. When shipped,
specimens should be packaged and labeled in compliance with applicable state, federal and international regulations covering the transport of
clinical specimens and infectious substances. Specimens should be shipped frozen.
 The sample volume required for a single determination of FT4 is 40 µL.
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WARNING AND PRECAUTIONS FOR USERS


 For In Vitro Diagnostic Use.
 Follow the package insert carefully. Reliability of assay results cannot be guaranteed if there are any deviations from the instructions in this
package insert.
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Safety Precautions
 CAUTION: This product requires the handling of human specimens. It is recommended that all human sourced materials be considered
potentially infectious and handled in accordance with the 29 CFR 1910.1030 Occupational exposure to bloodborne pathogens. Biosafety Level 2
or other appropriate biosafety practices should be used for materials that contain or are suspected of containing infectious agents.
 All samples, biological reagents and materials used in the assay should be considered potentially able to transmit infectious agents. They should
therefore be disposed of in accordance with the practices of your institution. Discard all materials in a safe and acceptable manner and in
compliance with prevailing regulatory requirements.
 This product contains Sodium Azide. Dispose of contents and containers must be in accordance with all local, regional and national regulations.
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 Refer to safety data sheets which are available on request.

Handling Precautions
 Do not use reagent kits beyond the expiration date.
 Do not interchange reagent components from different reagents or lots.
 Prior to loading the reagent kit on the system for the first time, the reagent kit requires mixing to re-suspend magnetic microbeads that have
settled during shipment.
 For magnetic microbeads mixing instructions, refer to the Preparation of the Reagent section of this package insert.
 To avoid contamination, wear clean gloves when operating with a reagent kit and sample.
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 Over time, residual liquids may dry on the septum surface. These are typically dried salts which have no effect on assay efficacy.
 For detailed discussion of handling precautions during system operation, refer to the SNIBE service information.

STORAGE AND STABILITY

 Sealed: Stored at 2-8°C until the expiration date.
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 Opened at 2-8°C: Minimum stability is 4 weeks.

 On-board: Minimum stability is 4 weeks.
 To ensure the best kit performance, it is recommended to place opened kits in the refrigerator after the end of the intraday test work. It is still
possible to keep on using the kit beyond the opened or on-board period if the controls are found within the expected ranges.
Keep upright for storage to facilitate later proper resuspension of magnetic microbeads.
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
 Keep away from sunlight.

TEST PROCEDURE
Preparation of the Reagent
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 Resuspension of the magnetic microbeads takes place automatically when the kit is loaded successfully, ensuring the magnetic microbeads are
totally resuspended homogenous prior to use.
 To ensure proper test performance, strictly adhere to the operating instructions of MAGLUMI series Fully-auto chemiluminescence immunoassay
analyzer. Each test parameter is identified via a RFID CHIP on the Reagent. For further information please refer to the operating instructions of
MAGLUMI series Fully-auto chemiluminescence immunoassay analyzer.

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 Please only refer to the current product lot insert enclosed with the kits package for execution and reporting

DILUTION
Samples for FT4 determinations cannot be diluted, as T4 in the blood is present in free and protein-bound forms which are in equilibrium. A
change in the concentration of the binding proteins alters this equilibrium.

LIMITATIONS
 Bacterial contamination or heat inactivation of the specimens may affect the test results.
 A result within the expected range does not rule out the presence of disease and should be interpreted together with other diagnostic procedures.
 Test results are reported quantitatively. However, diagnosis of a disease should not be based on the result of a single test, but should be
determined in conjunction with clinical findings in association with medical judgement.
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 Any therapeutical decision should also be taken on a case-by-case basis.
 Patient samples containing human anti-mouse antibodies (HAMA) may give falsely elevated or decreased values. Although HAMA-neutralizing
agents are added, extremely high HAMA serum concentrations may occasionally influence results.

RESULTS
Calculation of Results
The analyzer automatically calculates the FT4 concentration in each sample by means of a calibration curve which is generated by a 2-point calibration
master curve procedure. The results are expressed in pg/mL. For further information please refer to the operating instructions of MAGLUMI series
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Fully-auto chemiluminescence immunoassay analyzer.

Conversion factor: pg/mLx1.287=pmol/L
Interpretation of Results
The expected range for the FT4 assay was obtained by testing 292 apparently healthy individuals in China, and gave the following reference interval
listed below:
8.9-17.2 pg/mL (2.5th-97.5th percentiles)
Results may differ between laboratories due to variations in population and test method. It is recommended that each laboratory should establish its
own expected ranges.
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PERFORMANCE CHARACTERISTICS
Precision
Precision for the FT4 assay was determined as described in the CLSI EP5-A2. 3 controls and 3 human serum pools containing different
concentration of analyte were assayed in duplicate at two independent runs per day for 20 testing days. The result is summarized in the following
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table:
Mean(pg/mL) Within-Run Between-Run Total
Sample
(N=80) SD(pg/mL) %CV SD(pg/mL) %CV SD(pg/mL) %CV
Serum Pool 1 15.437 0.655 4.24 0.905 5.86 1.117 7.24
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Serum Pool 2 29.863 1.081 3.62 0.853 2.86 1.377 4.61

Serum Pool 3 79.805 2.201 2.76 1.206 1.51 2.510 3.15
Control 1 9.740 0.486 4.99 0.601 6.17 0.773 7.94
Control 2 20.944 0.881 4.21 0.708 3.38 1.130 5.40
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Control 3 45.958 1.353 2.94 1.681 3.66 2.158 4.70

Limit of Blank (LoB)

The LoB for the FT4 assay is 1.0 pg/mL.

Limit of Detection (LoD)

The LoD for the FT4 assay is 1.5 pg/mL.
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Measuring Range
1.0-120 pg/mL (defined by the limit of blank and the maximum of the master curve). Values below the limit of blank are reported as <1.0 pg/mL.
Values above the measuring range are reported as >120 pg/mL.

Method Comparison
A total of 100 clinical samples in the range of 3.85 to 105.20 pg/mL were tested using the FT4 assay (y) and a commercially available immunoassay (x)
according to CLSI EP09-A3. The data from the resulting linear regressions are summarized as:
y=1.019x -0.442, r2=0.997.
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Analytical Specificity
The Cross-reactivity of the the FT4 assay with a cross reactant can be expressed as the ratio of

 The amount of T4 required to displace 50% of the maximally bound labeled T4 from the anti-T4 antibody, and
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 The amount of the cross-reactant to give the same 50% displacement.

The results were listed in the following table:
Cross Reactant %Cross Reactivity
L-Triiodothyronine <0.017
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Diiodotyrosine <0.01
Monoiodotyrosine <0.01
3,5-Diiodo-L-thyronine <0.01
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Drugs up to the following concentrations did not interfere with the assay:
 Phenylbutazone 15.0 mg/dL
 Sodium salicylate 50.0 mg/dL
 Aspirin 50.0 mg/dL
 Ibuprofen 50.0 mg/dL
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 Acetaminophen 20.0 mg/dL

 Phenytoin 5.0 mg/dL
 Amiodarone 20.0 mg/dL
 Propylthiouracil 30.0 mg/dL

Endogenous Interference
Substances up to the following concentrations did not interfere with the assay:
 Bilirubin 41 mg/dL
 Triglyceride 2000 mg/dL
 Hemoglobin 2000 mg/dL
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 Total Protein 12 g/dL
 Rheumatoid factor 620 IU/mL
 HAMA 1232 ng/mL

REFERENCES
1. Robbins J, Rall JE. Interaction of thyroid hormones and protein in biological fluid. Recent ProgHorm Res 1957; 13:161-202.
2. Hay ID & Klee GG: Linking Medical Needs and Performance Goals: Clinical and Laboratory Perspectives on Thyroid Disease.
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ClinChem1993; 39 (7): 1519-1524.

3. Oppenheimer JH. Role of plasma proteins in the binding, distribution and metabolism of the thyroid hormones. N Engl J Med 1968;
278(21):1153-1162.
4. DeGroot LJ, Larsen PR, Refetoff S, Stanbury JB. Transport of Thyroid Hormone and Cell Uptake. In: The Thyroid and Its Diseases. New
York: Wiley and Sons, 1984; 62-65.
5. Hennemann G, Docter R, Krenning EP, Bos G, Otten M, Visser TJ. Raised Total Thyroxine and Free Thyroxine Index but Normal Free
Thyroxine. Lancet 1979; 1:639-642.
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Shenzhen New Industries Biomedical Engineering Co., Ltd.

No.16, Jinhui Road, Pingshan New District, Shenzhen, 518122, P.R. China
Tel: 0086-755-21536601 Fax:0086-755-28292740
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Lotus Global Co., Ltd.

1 Four Seasons Terrace, West Drayton, Middlesex London, UB7 9GG, United Kingdom
Tel: 0044-20-75868010 Fax: 0044-20-79006187

SYMBOLS EXPLANATIONS
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Consult instructions for use Manufacturer

Temperature limit
Use-by date
( Store at 2-8 °C)
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Contains sufficient for Keep away from sunlight

Authorized representative in the European

This way up
Community

In vitro diagnostic medical device Kit components

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Catalogue number Batch code

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