Anti-TSHR iFlash

Immunoassay Analyzer

Pretreating agent, phosphate buffer, 4.0

R3
REF C86049 2×50 Tests mL/bottle, with 0.05% ProClin 300.
Calibrator 1, 1 bottle, 1.0 mL, Tris buffer,
INTENDED USE with protein stabilizers and 0.05% ProClin
CAL1
The iFlash-Anti-TSHR assay is a paramagnetic particle 300. (See the concentration value in the
chemiluminescent immunoassay (CLIA) for the label)
quantitative determination of Anti-TSHR in human serum Calibrator 2, 1 bottle, 1.0 mL, Tris buffer,
using the iFlash Immunoassay Analyzer. with Anti-TSHR (mouse), protein stabilizers
CAL2
and 0.05% ProClin 300. (See the
SUMMARY AND EXPLANATION
concentration value in the label)
Autoimmune hyperthyreosis is caused by Anti-TSHR,
Calibrator 3, 1 bottle, 1.0 mL, Tris buffer,
which has similar stimulating thyroid function with TSH.
The stimulation of thyroid can be led by Anti-TSHR or TSH, with Anti-TSHR (mouse), protein stabilizers
CAL3
both of which can result in hyperthyreosis. Because and 0.05% ProClin 300. (See the
negative feedback system cannot control physiological concentration value in the label)
activities and metabolism of TSHR, this may always lead
to hyperthyreosis of Graves patients. Quantitative MATERIALS REQUIRED (BUT NOT PROVIDED)
determination of Anti-TSHR is of significance for it may not REF C89999/C89959/C89949, iFlash Pre-Trigger
only apply in diagnosis or removal of autoimmune Solution: hydrogen peroxide solution.
hyperthyreosis, but also monitor treatment and REF C89998/ C89958/ C89948, iFlash Trigger Solution:
palindromia of Graves disease patient. sodium hydroxide solution.

ASSAY PRINCIPLE REF C89997, iFlash Wash Buffer: phosphate buffered

saline solution with 0.05% ProClin 300.
The iFlash-Anti-TSHR assay is an indirect immunoassay.
REF C80001, iFlash Wash Buffer (10×): phosphate
 Incubation 1: have a reaction with sample, pretreating
buffered saline solution with 0.05% ProClin 300.
buffer solution and TSHR-receptor-coated
paramagnetic particles. Anti-TSHR in patient serum REF C89996, reaction vessels.
reacts with TSH receptor. Controls: Commercial controls could be used.
 Wash: the unbound materials are washed away from WARNINGS AND PRECAUTIONS
the solid phase in a magnetic field.
IVD For in vitro diagnostic use
 Incubation 2: add acridinium-ester-labeled second
 There is no testing method that can rule out the
antibody to react with antigen-antibody complex in
potential risk of infection with absolute certainty, so this
Incubation 1 and to form an antigen-antibody-second
product should be considered as potential biohazard.
antibody complex.
 Exercise the normal precautions required for handling
 Wash again.
all laboratory reagents.
 Trigger of signal: the Pre-Trigger and Trigger Solutions
 Disposal of all waste material should be in accordance
are added to the reaction mixture. The resulting
with local guidelines.
chemiluminescent reaction is measured as relative
light units (RLUs).  Wear gloves when handling specimens or reagents.
 A direct relationship exists between the amount of  Clean and disinfect all spills of specimens or reagents
Anti-TSHR in the sample and the RLUs detected by using a suitable disinfectant.
the iFlash optical system.  iFlash Trigger solution contains sodium hydroxide
 Results are determined via a calibration curve, which (NaOH) and should be avoided contact with eyes.
is instrument-specifically generated by 3-point REAGENT HANDLING
calibration and a master curve provided via the
 The reagents may not be used after the stated
reagent QR code.
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
TSHR-receptor (recombination protein) use. Close the bottles of calibrator right after
R1 coated paramagnetic particles, 3.5 calibration and store at 2–8°C.
mL/bottle, with 0.05% ProClin 300.
 Reagents of different bottles CANNOT be mixed with
Mouse anti human IgG each other.
R2 acridinium-ester-labeled marker, 4.0
 Prior to loading the iFlash-Anti-TSHR reagent on the
mL/bottle, with 0.05% ProClin 300. system for the first time, suspend the paramagnetic

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Anti-TSHR iFlash
Immunoassay Analyzer

particles by inverting the reagent bottle for 30 times code stuck on the inner face of reagent box. In case
slightly. the QR code cannot be read by barcode reader
 For further information on reagent handling automatically, enter the sequence number manually.
precautions during system operation, refer to the  Carry out calibration, if necessary.
iFlash system operating instruction.  Place the calibrators CAL1, CAL2, and CAL3 in the
calibrator rack in the sample zone. Only keep
STORAGE AND STABILITY
calibrators open during calibration.
Storage:  Test application.
 Store at 2–8°C in an upright position.  Load samples.
 The kit may be used immediately after removal from  Click RUN, the iFlash System performs all the
2-8°C storage. functions automatically and calculates the results.

Stability: CALIBRATION
 Unopened at 2–8°C: up to the stated expiration date.  Traceability: this method has been standardized
 Opened at 2–8°C: 28 days. against the NIBSC (National Institute for Biological
st
 Store on-board: 28 days. Standards and Control) 1 IS 90/672 Standard.
 Every iFlash-Anti-TSHR reagent kit has a QR code
SPECIMEN COLLECTION AND PREPARATION label containing the specific information for calibration
 Serum is the recommended sample. of the particular reagent lot.
 Ensure a complete clot formation of serum specimens  To perform an iFlash-Anti-TSHR calibration, test CAL1.
prior to centrifugation. (Clotting time should be no less CAL2, and CAL3 repeatedly, and the predefined
than 1 hour.) master curve is adapted to the analyzer.
 Centrifuge the specimens.  Once an iFlash Anti-TSHR calibration is accepted and
 Store specimens at room temperature (20 to 25°C) for stored, all subsequent samples may be tested without
no longer than 8 hours. further calibration unless:
 If the testing will not be completed within 2 hours,  A same reagent lot used over 28 days.
refrigerate the samples at 2 to 8°C.  Use a new lot of reagent kit.
 If the testing will not be completed within 3 days, or for  Control value is out of range.
shipment of samples, freeze at -20°C or colder in the  Required by pertinent regulations.
preservation of 12 months.
 Frozen specimens must be mixed thoroughly after MEASURING RANGE
thawing.  0.3 IU/L-40 IU/L

 The samples may be frozen for maximum 1 time. QUALITY CONTROL

 Only transfer the clarified specimen without the lipemic Use YHLO iFlash-Anti-TSHR Controls (with at least two
material from the centrifuged specimens with a lipid levels of concentration). Other suitable control materials
layer on the top into new sample tube. can be used.
 Ensure that residual fibrin and cellular matter have Quality control with at least two levels of concentration
been removed prior to analysis. should be run in every single test or every 24 hours after
 Be cautious when handling patient specimens to calibration.
prevent cross-contamination. Follow manufacturer’s instructions for reconstitution and
 Do not use heat-inactivated samples. storage. Establish target values and analytical ranges for
 Ensure the patient samples, calibrators and controls the control material based on the measurement
are at room temperature (20–25°C) before procedures in the lab. Results of quality control beyond
measurement. acceptable ranges may be invalid.
 Due to the possible evaporation, specimens and
RESULT
calibrators on the analyzers should be tested within 2
hours. Calculation:
The iFlash system automatically calculates the analyte
ASSAY PROCEDURE
concentration of each sample. The results are given in
 Refer to the system operating instruction or the online
IU/L.
help system for detailed information on preparing the
system. Expected Values:
 Specific information of the assay is stored in the QR It is recommended that each laboratory establish its own

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Anti-TSHR iFlash
Immunoassay Analyzer

expected reference range for the population of interest. Repetitiveness

Detection samples come from 150 cases of healthy Detect with repetitiveness reference in the concentration
population, 200 cases of non-Graves thyroid disease and of 5.0 IU/L (±20%) and 10.0 IU/L (±20%) respectively for
100 cases of untreated Graves patients to define the best 10 times. Coefficient of variation (CV) of repetitiveness
cut-off is 1.69 IU/L, sensitivity of this cut-off is 98.28% and (%CV): ≤ 10.0%.
specificity is 97.49%.
Analytical Specificity
LIMITATIONS Result of assay is less than 0.30 IU/L when human
 The iFlash-Anti-TSHR assay is limited to the thyroglobulin autoantibody <4,000 IU/mL;
determination of Anti-TSHR in human serum. Result of assay is less than 0.30 IU/L, when Anti-TPO
 The use of serum separator (gel) blood collection <600 IU/mL;
tubes has been validated in the use of the assay. Result of assay is less than 0.30 IU/L, when TSH <1,000
However, it is not possible to survey all manufacturers mIU/L.
or tube types.
Linearity
 If the results are inconsistent with clinical evidence,
Detect with linearity reference, correlation coefficient r≥
additional testing is suggested to confirm the result.
0.9900 in the range of 0.3 IU/L ~ 40 IU/L
 For diagnostic purposes, the results should be
interpreted in light of the total clinical presentation of Lot Tolerance
the patient, including symptoms, clinical history results. Use 3 lots of reagents to respectively detect repetitiveness
 Upper limit of measuring range of Anti-TSHR is 40 IU/L. in the concentration of 5.0 IU/L (±20%) and 10.0 IU/L
Samples over upper limit of measurement range may (±20%). Coefficient of variation (CV) of lot tolerance (%CV):
be diluted with sample diluent at the ratio of 1:5. ≤15.0%.
Diluted manually, the result should be multiplied by the
REFERENCES
dilution factor. The maximum dilution ratio should be
1. Pfannenstiel P, Saller B. Schilddrüsenkrankheiten –
no less than 1:10.
 Specimens from heparinized patients may be partially Diagnose und Therapie. Berliner Medizinische
coagulated and erroneous results could occur due to Verlagsanstalt,1995; 2: 28-30.
the presence of fibrin. 2. McIntosh RS, Asghar MS, Weetman AP. The
 The results from an alternative assays (i.e. EIA or RIA) antibody response in human autoimmune thyroid
may not be equivalent and cannot be used
disease. Clin Sci,1997; 92:529-541.
interchangeably.
 The assay is unaffected by icterus (bilirubin < 25 3. Costagliola S, Morgenthaler NG, Hoermann R, etal.
mg/dL), hemolysis (Hb < 0.4 g/dL), lipemia (Intralipid < Second generation assay for thyrotropin receptor
1,500 mg/dL), HAMA (< 600 ng/mL) and ANA (<500 antibodies has superior diagnostic sensitivity for
U/mL). Graves’ disease. J Clin Endocrinol Metab, 1999;
 No interference was observed from RF Effect up to a
84:90-97.
concentration of 600 IU/mL.
4. Quadbeck B, Hoermann R, Roggenbuck U, etal.
PERFORMANCE CHARACTERISTICS Sensitive thyrotropin and thyrotropin-receptor
Below are the representative performance data, and the
antibody determinations one month after
results obtained in individual laboratories may differ.
discontinuation of antithyroid drug treatment as
Precision predictors of relapse in Graves’ disease. Thyroid,
Detect with enterprise precision reference in the
concentration of 5.0 IU/L (±20%), 10.0 IU/L (±20%) and 2005; 15:1047-1054.
15.0 IU/L (±20%) respectively. Relative deviation of 5. Okamoto Y, Tanigawa SI, Ishikawa K, etal. TSH
precision should be in the range of ±10.0%. receptor antibody measurements and prediction of
remission in Graves’ disease patients treated with
Analytical Sensitivity
The detection limit representing the lowest measurable minimum maintenance doses of antithyroid drugs.
analyte level is 0.3 IU/L, which can be distinguished from Endocr J, 2006; 53(4):467-472.
zero. It is calculated as the value lying two standard
deviations above that of the lowest standard of the master
curve (standard 1 + 2 SD, n = 20).

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Anti-TSHR iFlash
Immunoassay Analyzer

SHENZHEN YHLO BIOTECH CO., LTD.

st th
1 -4 Floor, No.5 Building, Lishan Industrial
Area, Xinghai Road, Nanshan District,
Shenzhen 518054, P.R. China

Wellkang Ltd (www.CE-marking.eu)

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

ANNEX A:
Explanation of abbreviation

Abbreviation Explanation

Product No.

Calibrator

Reagent

Number of tests

Manufactured by

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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