T3 iFlash

Immunoassay Analyzer

REAGENTS
Reagent kit, 100 tests, 2 packs, 50 tests/pack
REF C86025 2×50 Tests
Streptavidin coated paramagnetic
R1 microparticles, 3.5 mL/pack, 0.05% ProClin
300.
INTENDED USE
The iFlash-T3 assay is a paramagnetic particle Anti-T3 acridinium-ester-labeled antibody; 4.0
R2
chemiluminescent immunoassay (CLIA) for the mL/pack; 0.05% ProClin 300.
quantitative determination of Total triiodothyronine (T3) in
human serum and plasma using the iFlash Immunoassay Biotin-labeled T3 conjugate,4.0 mL/pack;
R3
Analyzer. Phosphate buffer, 0.05% ProClin 300.

SUMMARY AND EXPLANATION Sample processing solution, 4.0mL/pack,

R4 8-anilino-1-naphthalenesulfonic acid
Triiodothyronine (T3) is a hormone that originates from magnesium, 0.05% ProClin 300.
direct thyroid synthesis and secretion (approximately 20%)
and from peripheral conversion of T4 toT3 (approximately Calibrator 1, 1 bottle, 1.0 mL, Phosphate
80%). T3 is secreted into the circulation in response to CAL1 buffer with protein stabilizers, 0.05% ProClin
TSH (thyroid stimulating hormone). The secretion of T3 is 300.
regulated by a negative feedback mechanism involving
Calibrator 2, 1 bottle, 1.0 mL, T3 in
the thyroid gland, pituitary gland, and hypothalamus.
CAL2 Phosphate buffer with protein stabilizers,
Primary malfunction of the thyroid gland may result in 0.05% ProClin 300.
excessive (hyper) or below normal (hypo) release of T3 or
T4. Disease related to thyroid-pituitary-hypothalamus Calibrator 3, 1 bottle, 1.0 mL, T3 in
system may influence the levels of T3 and T4. CAL3 Phosphate buffer with protein stabilizers,
0.05% ProClin 300.
The determination of T3 is utilized in the diagnosis of
T3-hyperthyroidism, the detection of early stages of Calibrator 4, 1 bottle, 1.0 mL, T3 in
hyperthyroidism and other thyroid diseases. CAL4 Phosphate buffer with protein stabilizers,
0.05% ProClin 300.
ASSAY PRINCIPLE
The iFlash-T3 assay is a competition immunoassay. MATERIALS REQUIRED (BUT NOT PROVIDED)
 First incubation: T3 in the sample and anti-T3 REF C89999/C89959/C89949, iFlash Pre-Trigger
acridinium-ester-labeled antibody react to form a Solution: hydrogen peroxide solution.
complex.
REF C89998/ C89958/ C89948, iFlash Trigger Solution:
 Second incubation: Add the biotin-labeled T3 sodium hydroxide solution.
conjugate and streptavidin coated paramagnetic
REF C89997, iFlash Wash Buffer: phosphate buffered
microparticles. After combining antibody and T3 in the
saline solution with 0.05% ProClin 300.
sample, the remaining site of antibody will react with
biotin-labeled T3 conjugate. REF C80001, iFlash Wash Buffer (10×): phosphate
buffered saline solution with 0.05% ProClin 300.
 Wash: The unbound materials are washed away from
the solid phase in a magnetic field. REF C89996, reaction vessels.
 Trigger of signal: The Pre-Trigger and Trigger Controls: Commercial controls could be used.
Solutions are added to the reaction mixture. The
WARNINGS AND PRECAUTIONS
resulting chemiluminescent reaction is measured as
IVD For in vitro diagnostic use
relative light units (RLUs).
 An inversely proportional relationship exists between  iFlash Trigger solution contains sodium hydroxide
the amount of T3 in the sample and the RLUs detected (NaOH) and should be avoided contact with eyes.
by the iFlash optical system.  Exercise the normal precautions required for handling
 Results are determined via a calibration curve, which all laboratory reagents.
is instrument-specifically generated by 4-point  Disposal of all waste material should be in accordance
calibration and a master curve provided via the with local guidelines.
reagent QR code.  Wear gloves when handling specimens or reagents.
 Clean and disinfect all spills of specimens or reagents
using a suitable disinfectant.

REAGENT HANDLING
 The reagents may not be used after the stated
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T3 iFlash
Immunoassay Analyzer

expiration date. before measurement.

 Avoid the formation of foam with all reagents.  Due to the possible evaporation, specimens and
 The reagents in the pack and calibrators are ready for calibrators on the analyzers should be measured
use. Close the bottles of calibrator right after within 2 hours.
calibration and store at 2 ~ 8°C.
ASSAY PROCEDURE
 Do not pool reagents within a reagent kit or between
 Refer to the system operating instruction or the online
reagent kits.
help system for detailed information on preparing the
 Prior to loading the iFlash-T3 reagent pack on the system.
system for the first time, resuspend the microparticles
 The test-specific parameters stored in barcode on the
by inverting the reagent pack slightly.
reagent pack are read in. In case the barcode cannot
 For further information on reagent handling be read, enter the sequence numbers.
precautions during system operation, refer to the
 Carry out calibration, if necessary.
iFlash system operating instruction.
 Place the calibrators CAL1, CAL2, CAL3 and CAL4in
STORAGE AND STABILITY the calibrator rack in the sample zone. Only keep
calibrators open during calibration.
Storage:
 Test application.
 Store at 2 ~ 8°C in an upright position.
 Loading samples (Use 80 μL of sample for each
 The kit may be used immediately after removal from 2 determination in addition to the sample container and
~ 8°C storage. 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. Alternate Result Unit

 Store on-board: 28 days.
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-T3 assay. Unit
 Ensure that serum specimens to form complete clot
prior to centrifugation. (Clotting time is not less than 1 ng/mL 1.536 nmol/L
hour).
 Centrifuge the specimens.
 Store specimens at room temperature (20 to 25°C) for CALIBRATION
no longer than 8 hours.  Traceability: This kit calibrator is traceable to the
 If the testing will not be completed within 8 hours, Chinese Academy of Food and Drug test T3 standard.
refrigerate the samples at 2 to 8°C.  Every iFlash-T3 reagent kit has a QR code label
 If the testing will not be completed within 3 days, or for containing the specific information for calibration of the
shipment of samples, freeze at -20°C or colder. particular reagent lot.

 Frozen specimens must be mixed thoroughly after  To perform an iFlash-T3 calibration, test CAL1, CAL2,
thawing. CAL3 and CAL4 in duplicate, and the predefined
master curve is adapted to the analyzer.
 The samples may be frozen for maximum1 time.
 Once an iFlash-T3 calibration is accepted and stored,
 Centrifuge specimens with a lipid layer on the top,
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
been removed prior to analysis.  A reagent kit with a new lot number is used.

 Use with caution in handling patient specimens to  Controls are out of range.
prevent cross-contamination.  Required by pertinent regulations.
 Do not use heat-inactivated samples. MEASURING RANGE
 Ensure that the patient samples, calibrators and 0.20 ~ 8.0 ng/mL
controls are at ambient temperature (20 ~ 25°C)
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T3 iFlash
Immunoassay Analyzer

QUALITY CONTROL  No interference was observed from HAMA up to a

Quality control materials should be run as single concentration of 600 ng/mL.
determinations at least once every 24 hours when the test
PERFORMANCE CHARACTERISTICS
is in use, once per reagent kit and after every calibration.
Include commercially available quality control materials Below are the representative performance data, and the
that cover at least two levels of analyte. Follow results obtained in individual laboratories may differ.
manufacturer’s instructions for reconstitution and storage. Precision
Each laboratory should establish mean values and
The precision of iFlash-T3 was determined using T3
acceptable ranges to assure proper performance. Quality
reagents and controls. Two controls, consisting low, and
control results that do not fall within acceptable ranges
median concentration of T3 were assayed.
may indicate invalid test results.
The within run precision was determined by testing each
RESULT sample in replicates of 10 (n = 10), and calculating percent
coefficient of variation (%CV). The results of the study are
Calculation:
shown below:
The iFlash system automatically calculates the analyte Sample Mean (ng/mL) SD %CV
concentration of each sample. The results are given in
ng/mL. 1 1.02 0.05 4.90

Expected Values: 2 3.95 0.19 4.81

A study of iFlash-T3 assay on samples from 364
apparently healthy people of various age groups yielded The between run precision was determined by testing
the following result: each sample in duplicate, two separate runs daily for 20
0.6 ~ 1.8 ng/mL. days (n = 80), and calculating percent coefficient of
variation (%CV). The results of the study are shown
It is recommended that each laboratory establish its own
below:
expected reference range for the specific population.
Sample Mean (ng/mL) SD %CV
LIMITATIONS
1 1.04 0.04 4.30
 The iFlash-T3 assay is limited to the determination of
T3 in human serum or plasma (lithium heparin, sodium 2 3.99 0.19 4.66
heparin, potassium EDTA, and sodium citrate). It has
not been validated for use with other types of plasma.
Analytical Sensitivity
 The use of serum separator (gel) blood collection
The detection limit representing the lowest measurable
tubes has been validated for use with this assay.
analyte level is 0.20ng/mL, which can be distinguished
However, it is not possible to survey all manufacturers
from zero. It is calculated as the value lying two standard
or tube types.
deviations above that of the lowest standard of the master
 The upper limit of the measuring range of this assay is curve (standard 1 + 2 SD, n = 20).
8.0 ng/mL. For the over-range samples, it is suggested
that using washing buffer 1:2 dilution. If using manual Method Comparison
dilution, the results should be multiplied by the dilution A comparison of the iFlash-T3 assay (y) with a
multiple. The biggest dilution ratio is not less than 1:5. commercially available T3 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=0.978x + 0.448
coagulated and erroneous results could occur due to r = 0.9940
the presence of fibrin. Sample concentration: 0.30 ~ 8.00 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) and total serum protein (< 10 g/dL). 1986; 305-318. Edited by Gornall, AG Philadelphia, PA:
 No interference was observed from rheumatoid factors J. B. Lippincott Co.
up to a concentration of 2,000 IU/mL. 2. White, GH. Recent advances in routine thyroid
 No interference was observed from anti-nuclear function testing. CRC - Critical reviews in clinical
antibodies up to a concentration of 500 U/mL. laboratory Sciences, 1986; 24: 315-362.
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T3 iFlash
Immunoassay Analyzer

3. Ekins, R. Measurement of free hormones in blood. ANNEX A:

Endocrinol. rev., 1986; 11:5-46. Explanation of abbreviation
4. Wilke, TJ. Estimation of free thyroid hormone
Abbreviation Explanation
concentrations in the clinical laboratory. Clinical
Chemistry. 1986; 32 (4): 585-592.
5. Guideline-Measurement of Free Thyroid Hormones, Product No.
C45A. 2004. National Committee for Clinical
Laboratory Standards. Calibrator

SHENZHEN YHLO BIOTECH CO., LTD. Reagent

1st-4th Floor, No.5 Building, Lishan Industrial

Area, Xinghai Road, Nanshan District, Number of tests
Shenzhen 518054, P.R. China
Manufactured by

Wellkang Ltd (www.CE-marking.eu)

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

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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