Anti-Tg iFlash

Immunoassay Analyzer

REAGENTS
REF C89003G 2×50 Tests Reagent kit, 100 tests, 2 packs, 50 tests/pack
Tg coated microparticles, 3.5 mL/pack,
R1
0.05% ProClin 300.
INTENDED USE
The iFlash-Anti-Tg assay is a paramagnetic particle Anti-Tg acridinium-ester-labeled conjugate,
R2
chemiluminescent immunoassay (CLIA) for the 6.5mL/pack; 0.05% ProClin 300.
quantitative determination of antibody to thyroglobulin (Tg)
Sample diluent, phosphate buffer, 10
in human serum and plasma using the iFlash R3
mL/pack; 0.05% ProClin 300.
Immunoassay Analyzer.
Calibrator 1, 1 bottle, 1.0 mL, phosphate
SUMMARY AND EXPLANATION CAL1 buffer with protein stabilizers, 0.05%
Thyroglobulin is synthesised in the thyrocytes, which ProClin 300.
release it into the follicular volume, where it plays an
important part in the production and storage of thyroid Calibrator 2, 1 bottle, 1.0 mL, Anti-Tg in
CAL2 phosphate buffer with protein stabilizers,
hormones. A concomitant elevation of autoantibodies to
0.05% ProClin 300.
both thyroid peroxidase (TPO) and Tg is indicative of a
hypertrophic form of autoimmune thyroiditis, namely Calibrator 3, 1 bottle, 1.0 mL, Anti-Tg in
lymphadenoid goitre (Hashimoto's disease), as well as of CAL3 phosphate buffer with protein stabilizers,
the atrophic form of autoimmune thyroiditis. 0.05% ProClin 300.

The measurement of autoantibodies against thyroglobulin

is useful in identifying patients with autoimmune thyroid MATERIALS REQUIRED (BUT NOT PROVIDED)
disease. Anti-Tg are associated with cases of mild REF C89999/C89959/C89949, iFlash Pre-Trigger
hypothyroidism or hyperthyroidism, and are frequently Solution: hydrogen peroxide solution.
found in patients with other autoimmune diseases such as REF C89998/ C89958/ C89948, iFlash Trigger Solution:
Rheumatoid Arthritis, Pernicious Anaemia and Type I sodium hydroxide solution.
Diabetes. Anti-Tg are detected in 30-60% of cases of
REF C89997, iFlash Wash Buffer: phosphate buffered
thyroid carcinoma patients. Furthermore, low levels of
saline solution with 0.05% ProClin 300.
Anti-Tg are also found in up to 20% of asymptomatic
individuals, particularly the elderly and more often in REF C80001, iFlash Wash Buffer (10×): phosphate
women than men, although the clinical significance of buffered saline solution with 0.05% ProClin 300.
these autoantibodies is unclear. REF C89996, reaction vessels.
Controls: Commercial controls could be used.
ASSAY PRINCIPLE
The iFlash-Anti-Tg assay is a sandwich immunoassay. WARNINGS AND PRECAUTIONS
 Incubation: Total Anti-Tg in the sample, Tg coated IVD For in vitro diagnostic use
paramagnetic microparticles and Anti-Tg  The calibrators (CAL2 and CAL3) have been prepared
acridinium-ester-labeled conjugate react to form a exclusively from the blood of donors tested individually
sandwich complex. and shown to be free from HBsAg, anti-HCV, and
 Wash: The unbound materials are washed away from anti-HIV-1/2 by approved methods.
the solid phase in a magnetic field.  However, no known test method can offer the
 Trigger of signal: The Pre-Trigger and Trigger complete assurance that products derived from human
Solutions are added to the reaction mixture. The sources will not transmit infection. Therefore, all
resulting chemiluminescent reaction is measured as humanized materials should be considered potentially
relative light units (RLUs). infectious.
 A direct relationship exists between the amount of total  Exercise the normal precautions required for handling
Anti-Tg in the sample and the RLUs detected by the all laboratory reagents.
iFlash optical system.  Disposal of all waste material should be in accordance
 Results are determined via a calibration curve, which with local guidelines.
is instrument-specifically generated by 3-point  Wear gloves when handling specimens or reagents.
calibration and a master curve provided via the  Clean and disinfect all spills of specimens or reagents
reagent QR code. using a suitable disinfectant.
 iFlash Trigger solution contains sodium hydroxide

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Anti-Tg iFlash
Immunoassay Analyzer

(NaOH) and should be avoided contact with eyes.  Ensure that residual fibrin and cellular matter have
been removed prior to analysis.
REAGENT HANDLING
 Use with caution in handling patient specimens to
 The reagents may not be used after the stated prevent cross-contamination.
expiration date.
 Do not use heat-inactivated samples.
 Avoid the formation of foam with all reagents.
 Ensure that the patient samples, calibrators and
 The reagents in the pack and calibrators are ready for controls are at ambient temperature (20–25°C) before
use measurement.
 Close the bottles of calibrator right after calibration and  Due to the possible evaporation, specimens and
store at 2–8°C. calibrators on the analyzers should be measured
 Do not pool reagents within a reagent kit or between within 2 hours.
reagent kits.
ASSAY PROCEDURE
 Prior to loading the iFlash-Anti-Tg reagent pack on the
system for the first time, suspend the microparticles by  Refer to the system operating instruction or the online
inverting the reagent pack slightly. help system for detailed information on preparing the
system.
 For further information on reagent handling
precautions during system operation, refer to the  The test-specific parameters stored in barcode on the
iFlash system operating instruction. reagent pack are read in. In case the barcode cannot
be read, enter the sequence numbers.
STORAGE AND STABILITY  Carry out calibration, if necessary.
Storage:  Place the calibrators CAL1, CAL2 and CAL3 in the
 Store at 2–8°C in an upright position. calibrator rack in the sample zone. Only keep
calibrators open during calibration.
 The kit may be used immediately after removal from
2-8°C storage.  Test application.
 Load samples (Use 20 μL of sample for each
Stability: determination in addition to the sample container and
 Unopened at 2–8°C: up to the stated expiration date. system dead volumes).
 Opened at 2–8°C: 28 days.  Click RUN, the iFlash System performs all the
 Store on-board: 28 days. functions automatically and calculates the results.

SPECIMEN COLLECTION AND PREPARATION CALIBRATION

 Serum or plasma (lithium heparin, sodium heparin  Traceability: This assay has been standardized
st
potassium EDTA, and sodium citrate) are the against the WHO 1 International Reference 65/093.
recommended samples. Other anticoagulants have  Every iFlash-Anti-Tg reagent kit has a QR code label
not been validated for use with the iFlash-Anti-Tg containing the specific information for calibration of the
assay. particular reagent lot.
 Ensure that serum specimens to form complete clot  To perform an iFlash-Anti-Tg calibration, test CAL1,
prior to centrifugation. CAL2 and CAL3 in duplicate, and the predefined
 Centrifuge the specimens. master curve is adapted to the analyzer.
 Store specimens at room temperature (20 to 25°C) for  Once an iFlash-Anti-Tg calibration is accepted and
no longer than 8 hours. stored, all subsequent samples may be tested without
 If the testing will not be completed within 8 hours, further calibration unless:
refrigerate the samples at 2 to 8°C.  After 28 days when using the same reagent lot.
 If the testing will not be completed within 14 days, or  A reagent kit with a new lot number is used.
for shipment of samples, freeze at -20°C or colder.  Controls are out of range.
 Frozen specimens must be mixed thoroughly after  Required by pertinent regulations.
thawing.
MEASURING RANGE
 The samples may be frozen for maximum 1 time.
 1 -1,000 IU/mL
 Centrifuge specimens with a lipid layer on the top, and
transfer only the clarified specimen without the lipemic QUALITY CONTROL
material. Quality control materials should be run as single
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Anti-Tg iFlash
Immunoassay Analyzer

determinations at least once every 24 hours when the test  The assay is unaffected by icterus (bilirubin < 10
is in use, once per reagent kit and after every calibration. mg/dL), hemolysis (Hb < 500 mg/dL), lipemia
Include commercially available quality control materials (Intralipid < 1,800 mg/dL) and total serum protein (< 10
that cover at least two levels of analyte. Follow g/dL).
manufacturer’s instructions for reconstitution and storage.  No interference was observed from rheumatoid factors
Each laboratory should establish mean values and up to a concentration of 2,000 IU/mL.
acceptable ranges to assure proper performance. Quality
 No interference was observed from anti-nuclear
control results that do not fall within acceptable ranges
antibodies up to a concentration of 500 AU/mL.
may indicate invalid test results.
 No interference was observed from HAMA up to a
RESULT concentration of 600 ng/mL.

Calculation: PERFORMANCE CHARACTERISTICS

The iFlash system automatically calculates the analyte Below are the representative performance data, and the
concentration of each sample. The results are given in results obtained in individual laboratories may differ
IU/mL
Precision
Interpretation of Results The precision of iFlash-Anti-Tg was determined using
A study of with iFlash-Anti-Tg assay on samples from 369 Anti-Tg reagents, samples and controls. Three serum
apparently healthy males of various age groups yielded samples, consisting low, median, and high concentration
the following result: of Anti-Tg were assayed.
< 114 IU/mL (95th percentile) The within run precision was determined by testing each
It is recommended that each laboratory establish its own sample in replicates of 10 (n = 10), and calculating percent
expected reference range for the specific population. coefficient of variation (%CV). The results of the study are
shown below:
LIMITATIONS Sample Mean (IU/mL) SD %CV
 The iFlash-Anti-Tg assay is limited to the 1 20.66 0.96 4.65%
determination of Anti-Tg in human serum or plasma 2 115.91 4.20 3.62%
(lithium heparin, sodium heparin, potassium EDTA,
3 199.45 10.14 5.08%
and sodium citrate). It has not been validated for use
with other types of plasma. The between run precision was determined by testing
 The use of serum separator (gel) blood collection each sample in duplicate, two separate runs daily for 20
tubes has been validated for use with this assay. days (n = 80), and calculating percent coefficient of
However, it is not possible to survey all manufacturers variation (%CV). The results of the study are shown
or tube types. below:
Sample Mean (IU/mL) SD %CV
 The upper limit of the measuring range of this assay is
1,000 IU/mL. Over-range samples may be diluted with 1 20.54 0.99 4.82%
wash buffer (1:20 recommended) and re-tested to 2 200.02 8.84 4.42%
obtain an estimate of the actual concentration.
Analytical Sensitivity
 If the results are inconsistent with clinical evidence,
additional testing is suggested to confirm the result. The detection limit representing the lowest measurable
analyte level is 1 IU/mL, which can be distinguished from
 For diagnostic purposes, the results should be
zero. It is calculated as the value lying two standard
interpreted in light of the total clinical presentation of
deviations above that of the lowest standard of the master
the patient, including symptoms, clinical history results.
curve (standard 1 + 2 SD, n = 20).
 Specimens from heparinized patients may be partially
coagulated and erroneous results could occur due to Method Comparison
the presence of fibrin. A comparison of the iFlash-Anti-Tg assay (y) with a
 The results from an alternative assays (i.e. EIA or RIA) commercially available Anti-Tg assay (x) using clinical
may not be equivalent and cannot be used samples was performed, and the curve is fitted with Linear
interchangeably. regression)
 Samples containing an apparent Anti-Tg level as high y = 1.001x + 41.85
as 100,000 IU/mL did not exhibit a hook effect in the r = 0.996
iFlash-Anti-Tg assay. Sample concentration: 1 – 1,000 IU/mL

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Anti-Tg iFlash
Immunoassay Analyzer

Number of samples measured: 88 ANNEX A:

Explanation of abbreviation
REFERENCES
1. Roitt IM, Doniach D, Campbell PN, et al. Abbreviation Explanation
Auto-antibodies in Hashimoto’s Disease
(Lymphadenoid Goitre). Lancet 1956; 6947:820-1. Product No.
2. Ruf J, Feldt-Rasmussen U, Heged s L, et al.
Bispecific thyroglobulin and thyroperoxidase
Calibrator
autoantibodies in patients with various thyroid and
autoimmune diseases. J Clin Endocrinol Metab 1994;
79(5): 1404-9. Reagent
3. Scherbaum WA. On the clinical importance of thyroid
microsomal and thyroglobulin antibody determination. Number of tests
Acta Endocrinol (Copenh) 1987; S281: 325-9.
4. Walker DJ, Griffiths M, Griffiths ID. Occurence of Manufactured by
autoimmune diseases and autoantibodies in
multicase rheumatoid arthritis families. Ann Rheum
Dis 1986; 45: 323-6. EU Representative

5. Feldt-Rasmussen U, Rasmussen K. Serum

thyroglobulin (Tg) in presence of thyroglobulin EC Declaration of Conformity
autoantibodies (TgAb). Clinical and methodological
relevance of the interaction between Tg and TgAb in Caution
vitro and in vivo. J Endocrinol Invest 1985; 8: 571-6.
6. Schaadt B, Feldt-Rasmussen U, Rasmusson B, et al.
Instructions for use
Assessment of the influence of thyroglobulin (Tg)
autoantibodies and other interfering factors on the In vitro diagnostic medical
use of serum Tg as tumor marker in differentiated device
thyroid carcinoma. Thyroid 1995; 5(3): 165-70.
7. Ericsson U-B, Christensen SB, Thorell JI. A high Lot No.
prevalence of thyroglobulin autoantibodies in adults
with and without thyroid disease as measured with a Date of manufacture
sensitive solid-phase immunosorbent radioassay.
Clin Immunol Immunopathol 1985; 37: 154-62.
Expiry date
8. Weetman AP and McGregor AM. Autoimmune thyroid
disease: further developments in our understanding.
Endocrine Reviews 1994; 15(6): 788-830. Biohazard Symbol

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