T4 iFlash

Immunoassay Analyzer

T4 acridinium-ester-labeled conjugate, 4.0

REF C86026 R2
2×50 Tests mL/pack, 0.05% ProClin 300.

Sample processing solution, 4.0 mL/pack,

INTENDED USE R3 8-anilino-1-naphthalenesulfonic acid
magnesium, 0.05% ProClin 300.
The iFlash-T4 assay is a paramagnetic particle
chemiluminescent immunoassay (CLIA) for the Calibrator 1, 1 bottle, 1.0 mL, Phosphate
quantitative determination of Total thyroxine (T4) in human CAL1 buffer with protein stabilizers, 0.05% ProClin
serum and plasma using the iFlash Immunoassay 300.
Analyzer.
Calibrator 2, 1 bottle, 1.0 mL, T4 in
SUMMARY AND EXPLANATION CAL2 Phosphate buffer with protein stabilizers,
Thyroxine (T4) is a hormone synthesized and secreted by 0.05% ProClin 300.
the thyroid gland, and plays an important role in regulating
Calibrator 3, 1 bottle, 1.0 mL, T4 in
metabolism system. T4 is secreted into the circulation in CAL3 Phosphate buffer with protein stabilizers,
response to the pituitary hormone TSH (thyroid stimulating 0.05% ProClin 300.
hormone). T4 and its associate thyroid hormone T3 are
responsible for regulating diverse biochemical processes Calibrator 4, 1 bottle, 1.0 mL, T4 in
throughout the body that are essential for normal CAL4 Phosphate buffer with protein stabilizers,
metabolic and neural activity. 0.05% ProClin 300.

The determination of T4 can be utilized for the following

indications: the detection of hyperthyroidism, the detection MATERIALS REQUIRED (BUT NOT PROVIDED)
of primary and secondary hypothyroidism and the REF C89999/C89959/C89949, iFlash Pre-Trigger
monitoring of TSH-suppression therapy. Solution: hydrogen peroxide solution.
REF C89998/ C89958/ C89948, iFlash Trigger Solution:
ASSAY PRINCIPLE
sodium hydroxide solution.
The iFlash-T4 assay is a competitive immunoassay.
REF C89997, iFlash Wash Buffer: phosphate buffered
 First incubation: T4 in the sample react with sample
saline solution with 0.05% ProClin 300.
processing solution, T4 is released.
REF C80001, iFlash Wash Buffer (10×): phosphate
 Second incubation: Add the acridinium-ester-labeled
buffered saline solution with 0.05% ProClin 300.
T4 conjugate and anti-T4 antibody coated
REF C89996, reaction vessels.
paramagnetic microparticles. T4 in the sample and the
acridinium-ester-labeled T4 conjugate can react with Controls: Commercial controls could be used.
anti-T4 antibody competitively, to form a complex of
WARNINGS AND PRECAUTIONS
antibody and acridinium-ester-labeled antigen.
IVD For in vitro diagnostic use
 Wash: The unbound materials are washed away from
 iFlash Trigger solution contains sodium hydroxide
the solid phase in a magnetic field.
(NaOH) and should be avoided contact with eyes.
 Trigger of signal: The Pre-Trigger and Trigger
 Exercise the normal precautions required for handling
Solutions are added to the reaction mixture. The
all laboratory reagents.
resulting chemiluminescent reaction is measured as
relative light units (RLUs).  Disposal of all waste material should be in accordance
with local guidelines.
 An inversely proportional relationship exists between
the amount of T4 in the sample and the RLUs detected  Wear gloves when handling specimens or reagents.
by the iFlash optical system.  Clean and disinfect all spills of specimens or reagents
 Results are determined via a calibration curve, which using a suitable disinfectant.
is instrument-specifically generated by 4-point
REAGENT HANDLING
calibration and a master curve provided via the
reagent QR code.  The reagents may not be used after the stated
expiration date.
REAGENTS  Avoid the formation of foam with all reagents.
Reagent kit, 100 tests, 2 packs, 50 tests/pack  The reagents in the pack and calibrators are ready for
use. Close the bottles of calibrator right after
Anti-T4 antibody coated paramagnetic
R1 microparticles, 3.5 mL/pack, 0.05% ProClin calibration and store at 2 ~ 8°C.
300.  Do not pool reagents within a reagent kit or between
reagent kits.

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T4 iFlash
Immunoassay Analyzer

 Prior to loading the iFlash-T4 reagent pack on the help system for detailed information on preparing the
system for the first time, resuspend the microparticles system.
by inverting the reagent pack slightly.  The test-specific parameters stored in barcode on the
 For further information on reagent handling reagent pack are read in. In case the barcode cannot
precautions during system operation, refer to the be read, enter the sequence numbers.
iFlash system operating instruction.  Carry out calibration, if necessary.
STORAGE AND STABILITY  Place the calibrators CAL1, CAL2, CAL3 and CAL4in
the calibrator rack in the sample zone. Only keep
Storage: calibrators open during calibration.
 Store at 2–8°C in an upright position.  Test application.
 The kit may be used immediately after removal from 2  Loading samples (Use 20 μL of sample for each
~ 8°C storage. determination in addition to the sample container and
system dead volumes).
Stability:
 Click RUN, the iFlash System performs all the
 Unopened at 2 ~ 8°C: up to the stated expiration date.
functions automatically and calculates the results.
 Opened at 2 ~ 8°C: 28 days.
 Store on-board: 28 days. Alternate Result Unit
Conversion formula:
SPECIMEN COLLECTION AND PREPARATION
(Concentration in YHLO Unit) x (Conversion Factor) =
 Serum or plasma (lithium heparin, sodium heparin
(Concentration in Standard Unit)
potassium EDTA, and sodium citrate) are the
recommended samples. Other anticoagulants have YHLO Unit Conversion Factor Standard
not been validated for use with the iFlash-T4 assay. Unit
 Ensure that serum specimens to form complete clot
prior to centrifugation. (Clotting time is not less than 1 ng/mL 1.287 nmol/L
hour).
 Centrifuge the specimens.
CALIBRATION
 Store specimens at room temperature (20 to 25°C) for
no longer than 8 hours.  Traceability: This kit calibrator is traceable to the
Chinese Academy of Food and Drug test T4 standard.
 If the testing will not be completed within 8 hours,
refrigerate the samples at 2 to 8°C.  Every iFlash-T4 reagent kit has a QR code label
containing the specific information for calibration of the
 If the testing will not be completed within 3 days, or for
particular reagent lot.
shipment of samples, freeze at -20°C or colder.
 To perform an iFlash-T4 calibration, test CAL1, CAL2,
 Frozen specimens must be mixed thoroughly after
CAL3 and CAL4 in duplicate, and the predefined
thawing.
master curve is adapted to the analyzer.
 The samples may be frozen for maximum 1 time.
 Once an iFlash-T4 calibration is accepted and stored,
 Centrifuge specimens with a lipid layer on the top, and all subsequent samples may be tested without further
transfer only the clarified specimen without the lipemic calibration unless:
material.
 After 28 days when using the same reagent lot.
 Ensure that residual fibrin and cellular matter have
 A reagent kit with a new lot number is used.
been removed prior to analysis.
 Controls are out of range.
 Use with caution in handling patient specimens to
prevent cross-contamination.  Required by pertinent regulations.
 Do not use heat-inactivated samples. MEASURING RANGE
 Ensure that the patient samples, calibrators and  5 ~ 300 ng/mL
controls are at ambient temperature (20 ~ 25°C)
before measurement. QUALITY CONTROL
 Due to the possible evaporation, specimens and Quality control materials should be run as single
calibrators on the analyzers should be measured determinations at least once every 24 hours when the test
within 2 hours. is in use, once per reagent kit and after every calibration.
Include commercially available quality control materials
ASSAY PROCEDURE that cover at least two levels of analyte. Follow
 Refer to the system operating instruction or the online manufacturer’s instructions for reconstitution and storage.

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T4 iFlash
Immunoassay Analyzer

Each laboratory should establish mean values and Precision

acceptable ranges to assure proper performance. Quality The precision of iFlash-T4 was determined using T4
control results that do not fall within acceptable ranges reagents and controls. Two controls, consisting low, and
may indicate invalid test results. median concentration of T4 were assayed.
RESULT The within run precision was determined by testing each
sample in replicates of 10 (n = 10), and calculating percent
Calculation: coefficient of variation (%CV). The results of the study are
The iFlash system automatically calculates the analyte shown below:
concentration of each sample. The results are given in Sample Mean (ng/mL) SD %CV
ng/mL.
1 18.72 0.89 4.76
Expected Values:
A study of iFlash-T4 assay on samples from 295 2 99.13 4.61 4.65
apparently healthy people of various age groups yielded
The between run precision was determined by testing
the following result:
th th each sample in duplicate, two separate runs daily for 20
45 ~ 126 ng /mL (2.5 ~ 97.5 percentile)
days (n = 80), and calculating percent coefficient of
It is recommended that each laboratory establish its own variation (%CV). The results of the study are shown
expected reference range for the specific population. below:

LIMITATIONS Sample Mean (ng/mL) SD %CV

 The iFlash-T4 assay is limited to the determination of 1 18.25 0.79 4.35

T3 in human serum or plasma (lithium heparin, sodium
heparin, potassium EDTA, and sodium citrate). It has 2 100.44 4.93 4.91
not been validated for use with other types of plasma.
 The use of serum separator (gel) blood collection Analytical Sensitivity
tubes has been validated for use with this assay. The detection limit representing the lowest measurable
However, it is not possible to survey all manufacturers analyte level is 5 ng/mL, which can be distinguished from
or tube types. zero. It is calculated as the value lying two standard
 The upper limit of the measuring range of this assay is deviations above that of the lowest standard of the master
300 ng/mL. For the over-range samples, it is curve (standard 1 + 2 SD, n = 20).
suggested that using washing buffer 1:2 dilution. If
using manual dilution, the results should be multiplied Method Comparison
by the dilution multiple. The biggest dilution ratio is not A comparison of the iFlash-T4 assay (y) with a
less than 1:5. commercially available T4 assay (x) using clinical samples
 If the results are inconsistent with clinical evidence, was performed, and the curve is fitted with Linear
additional testing is suggested to confirm the result. regression)
 Specimens from heparinized patients may be partially y = 1.013x +9.690
coagulated and erroneous results could occur due to r = 0.9945
the presence of fibrin. Sample concentration: 6.00 ~ 300 ng/mL
 The results from an alternative assays (i.e. EIA or RIA) Number of samples measured: 90
may not be equivalent and cannot be used
interchangeably. REFERENCES
 The assay is unaffected by icterus (bilirubin < 10 1. Gornall, AG, Luxton, AW, Bhavnani, BR. Endocrine
mg/dL), hemolysis (Hb < 500 mg/dL), lipemia (Intralipid disorders in applied biochemistry of clinical disorders,
< 1,800 mg/dL). 305–318. Edited by Gornall, AG Philadelphia, PA: J B
Lippincott Co, 1986.
 No interference was observed from rheumatoid factors
up to a concentration of 2,400 IU/mL. 2. White, GH, Recent advances in routine thyroid
function testing, CRC – Critical Reviews in Clinical
 No interference was observed from anti-nuclear
Laboratory Sciences, 24: 315-362: 1987.
antibodies up to a concentration of 500 U/mL.
3. Wilke, TJ. Estimation of free thyroid hormone
 No interference was observed from HAMA up to a
concentrations in the clinical laboratory. Clinical
concentration of 600 ng/mL.
Chemistry. 1986;32 (4): 585-592.
PERFORMANCE CHARACTERISTICS 4. Spencer, CA. Thyroid status: trends in testing –
Below are the representative performance data, and the selective test use cuts cost. Clinical Chemistry News.
results obtained in individual laboratories may differ. November,1989; 15 (11): 9-14.
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T4 iFlash
Immunoassay Analyzer

5. Liewendahl, K, Majuri, H, Helenius, T. Thyroid function ANNEX A:

tests in patients on long-term treatment with various Explanation of abbreviation
anticonvulsant drugs. Clinical Endocrinology, 1978; 8:
187-191. Abbreviation Explanation
6. Wenzel, KW. Pharmacological interference with in vitro
tests of thyroid function. Metabolis. 1981; 30 (7): Product No.
717-732.
Calibrator
SHENZHEN YHLO BIOTECH CO., LTD.
Reagent
1st-4th Floor, No.5 Building, Lishan Industrial
Area, Xinghai Road, Nanshan District,
Shenzhen 518054, P.R. China Number of tests

Wellkang Ltd (www.CE-marking.eu) Manufactured by

Suite B, 29 Harley St., London W1G 9QR, UK

EU Representative

EC Declaration of Conformity

Caution

Instructions for use

In vitro diagnostic medical

device

Lot No.

Date of manufacture

Expiry date

Biohazard Symbol

Pictograms for Caution

Pictograms for Hazardous to the

aquatic environment

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