05572339001V3

Anti-TPO
Antibodies to thyroid peroxidase

REF 11820818 191 100 tests Reagents - working solutions

• Indicates analyzers on which the kit can be used M Streptavidin-coated microparticles (transparent cap), 1 bottle, 6.5 mL:
Streptavidin-coated microparticles 0.72 mg/mL; preservative.
MODULAR R1 Anti-TPO-Ab~Ru(bpy)2+ 3 (gray cap), 1 bottle, 10 mL:
Elecsys 2010 ANALYTICS cobas e 411 cobas e 601 cobas e 602 Polyclonal anti-TPO antibody (sheep) labeled with ruthenium complex
E170 1.0 mg/L; TRIS buffer 100 mmol/L, pH 7.2; preservative.
• • • • • R2 TPO~biotin (black cap), 1 bottle, 10 mL:
Biotinylated TPO (recombinant) 0.15 mg/L; TRIS buffer
English 30 mmol/L, pH 7.0; preservative.
Cal1 A-TPO calibrator 1 (white cap), 1 bottle, 1.5 mL:
Intended use Polyclonal anti-TPO antibody (sheep) approx. 35 IU/mL in
Immunoassay for the in vitro quantitative determination of antibodies to thyroid human serum matrix; preservative.
peroxidase in human serum and plasma. The anti-TPO determination is Cal2 A-TPO calibrator 2 (black cap), 1 bottle, 1.5 mL:
used as an aid in the diagnosis of autoimmune thyroid diseases. Polyclonal anti-TPO antibody (sheep) approx. 350 IU/mL in
The electrochemiluminescence immunoassay “ECLIA” is intended for human serum matrix; preservative.
use on Elecsys and cobas e immunoassay analyzers. Calibrators: The exact lot-specific calibrator values are encoded in
Summary1,2,3,4,5,6 the barcoded labels of the test-specific reagent.
Thyroid-specific peroxidase (TPO) is present on the microsomes of thyrocytes Precautions and warnings
and is expressed at its apical cell surface. In synergy with thyroglobulin For in vitro diagnostic use.
(Tg) this enzyme has an essential function in the iodination of L-tyrosine Exercise the normal precautions required for handling all laboratory reagents.
and the chemical coupling of the resulting mono- and di-iodotyrosine Disposal of all waste material should be in accordance with local guidelines.
to form the thyroid hormones T4, T3, and rT3. Safety data sheet available for professional user on request.
TPO is a potential autoantigen. Elevated serum titers of antibodies to TPO are All human material should be considered potentially infectious.
found in several forms of thyroiditis caused by autoimmunity. The still frequently All products derived from human blood are prepared exclusively
found term “microsomal antibody” originates from the time when TPO had not from the blood of donors tested individually and shown to be free
yet been identified as an antigen in autoimmunity caused by microsomes. In the from HBsAg and antibodies to HCV and HIV.
clinical sense the two terms anti-TPO and microsomal antibody can be used The testing methods applied were FDA-approved or cleared in compliance
synonymously; there are differences, however, with regard to the test methods. with the European Directive 98/79/EC, Annex II, List A.
High anti-TPO titers are found in up to 90 % of patients with chronic The materials of human origin used for the controls were tested for HIV,
Hashimoto’s thyroiditis. In Graves’ disease, 70 % of the patients have HBV, and HCV infection. The findings were negative.
an elevated titer. Although the sensitivity of the procedure can be However, as no testing method can rule out the potential risk of infection
increased by simultaneously determining other thyroid antibodies (anti-Tg, with absolute certainty, the material should be treated just as carefully
TSH-receptor-antibody - TRAb), a negative finding does not rule out the as a patient specimen. In the event of exposure the directives of the
possibility of an autoimmune disease. The magnitude of the antibody titer does responsible health authorities should be followed.7,8
not correlate with the clinical activity of the disease. Initially elevated titers Avoid the formation of foam with all reagents and sample types
can become negative after lengthy periods of illness or during remission. If (specimens, calibrators, and controls).
antibodies reappear following remission, then a relapse is probable.
Whereas the usual microsomal antibody tests employ unpurified microsomes Reagent handling
as an antigen preparation, the anti-TPO tests use a purified peroxidase. The The reagents in the kit are ready for use and are supplied in
two procedures are of comparable performance in terms of clinical sensitivity, bottles compatible with the system.
but better lot-to-lot consistency and higher clinical specificity can be expected Elecsys 2010 and cobas e 411 analyzers: The Elecsys calibrators Cal1 and
from anti-TPO tests due to the higher quality of the antigen used. Cal2 should only be left on the analyzers at 20-25 °C during calibration. After
Recombinant antigen and polyclonal anti-TPO antibodies are use, close the bottles as soon as possible and store at 2-8 °C.
used in the Elecsys Anti-TPO assay. Ensure that no calibration solution is trapped in the opened snap-cap.
Test principle Because of possible evaporation effects, not more than 5 calibration
Competition principle. Total duration of assay: 18 minutes. procedures per bottle set should be performed.
• 1st incubation: 20 µL of sample are incubated with anti-TPO-antibodies MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 analyzers:
labeled with a ruthenium complexa. Unless the entire volume is necessary for calibration on the analyzer, transfer
• 2nd incubation: After addition of biotinylated TPO and streptavidin-coated aliquots of the ready-for-use calibrators into empty snap-cap bottles (CalSet
microparticles, the anti-TPO antibodies in the sample compete Vials). Attach the supplied labels to these additional bottles. Store the aliquots
with the ruthenium-labeled anti-TPO antibodies for the biotinylated for later use at 2-8 °C. Perform only one calibration procedure per aliquot.
TPO antigen. The entire complex becomes bound to the solid All information required for correct operation is read in via the
phase via interaction of biotin and streptavidin. respective reagent barcodes.
• The reaction mixture is aspirated into the measuring cell where the Storage and stability
microparticles are magnetically captured onto the surface of the Store at 2-8 °C.
electrode. Unbound substances are then removed with ProCell. Store the Elecsys Anti-TPO reagent kit (M, R1, R2) upright in order to ensure
Application of a voltage to the electrode then induces chemiluminescent complete availability of the microparticles during automatic mixing prior to use.
emission which is measured by a photomultiplier. Stability:
• Results are determined via a calibration curve which is unopened at 2-8 °C up to the stated expiration date
instrument-specifically generated by 2-point calibration and a
after opening at 2-8 °C 6 weeks
master curve provided via the reagent barcode.
on the analyzers 2 weeks
a) Tris(2,2’-bipyridyl)ruthenium(II)-complex (Ru(bpy)2+
3 )
Cal1, Cal2 after opening 6 weeks at 2-8 °C
on Elecsys 2010 and cobas e 411 at up to 5 hours
20-25 °C
on MODULAR ANALYTICS E170, use only once
cobas e 601 and cobas e 602

2010-10, V 3 English 1/3 Elecsys and cobas e analyzers

Anti-TPO
Antibodies to thyroid peroxidase
Store the calibrators upright! Do not freeze! Ensure that no calibration Resuspension of the microparticles takes place automatically before use.
solution is trapped in the opened snap-cap. Read in the test-specific parameters via the reagent barcode. If in exceptional
cases the barcode cannot be read, enter the 15-digit sequence of numbers.
Specimen collection and preparation
Bring the cooled reagents (M, R1, R2) to approx. 20 °C and place
Only the specimens listed below were tested and found acceptable.
on the reagent disk (20 °C) of the analyzer. Avoid the formation of
Serum collected using standard sampling tubes or tubes
foam. The system automatically regulates the temperature of the
containing separating gel.
reagents and the opening/closing of the bottles.
Li-, Na-, NH+4-heparin, K3-EDTA, sodium citrate, and sodium
fluoride/potassium oxalate plasma. Place the Elecsys calibrators A-TPO Cal1 and Cal2 in the sample zone of the
Criterion: Recovery within 90-110 % of serum value or slope 0.9-1.1 + intercept analyzer. Only keep open during calibration. All information necessary for
within < ± 2 x analytical sensitivity (LDL) + coefficient of correlation > 0.95. calibration is encoded on the barcoded bottle labels and is read in automatically.
Stable for 3 days at 2-8 °C, at least 1 month at -20 °C. Freeze only once.9 After calibration has been performed, store Cal1 and Cal2 at 2-8 °C or discard
The sample types listed were tested with a selection of sample collection tubes (MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 analyzers).
that were commercially available at the time of testing, i.e. not all available Calibration
tubes of all manufacturers were tested. Sample collection systems from Traceability: This method has been standardized against the NIBSC (National
various manufacturers may contain differing materials which could affect Institute for Biological Standards and Control) 66/387 Standard.
the test results in some cases. When processing samples in primary tubes Every Elecsys Anti-TPO reagent set (M, R1, R2) has a barcoded
(sample collection systems), follow the instructions of the tube manufacturer. label containing the specific information for calibration of the particular
Centrifuge samples containing precipitates before performing the reagent lot. The predefined master curve is adapted to the analyzer
assay. Do not use heat-inactivated samples. Do not use samples by the use of Elecsys Anti-TPO Cal1 and Cal2.
and controls stabilized with azide. Calibration frequency: Perform calibration on all analyzers as follows:
Ensure the patients’ samples, calibrators, and controls are at ambient
• with every reagent kit
temperature (20-25 °C) before measurement.
Because of possible evaporation effects, samples, calibrators, and controls Renewed calibration on all analyzers:
on the analyzers should be measured within 2 hours. • daily: when using the same reagent kit on the analyzers
• as required: e.g. quality control findings outside the specified limits
Materials provided
See “Reagents - working solutions” section for reagents. Quality control
• 2 x 6 bottle labels For quality control, use Elecsys PreciControl ThyroAB 1 and 2.
Other suitable control material can be used in addition.
Materials required (but not provided) Controls for the various concentration ranges should be run as single
• REF 05042666191, PreciControl ThyroAB, for 2 x 2 mL each determinations at least once every 24 hours when the test is in use, once
of PreciControl ThyroAB 1 and 2 per reagent kit, and after every calibration. The control intervals and
limits should be adapted to each laboratory’s individual requirements.
• REF 11776576322, CalSet Vials, 2 x 56 empty snap-cap bottles
Values obtained should fall within the defined limits.
• REF 11732277122, Diluent Universal, 2 x 16 mL sample diluent or Each laboratory should establish corrective measures to be taken
REF 03183971122, Diluent Universal, 2 x 36 mL sample diluent if values fall outside the limits.
• General laboratory equipment Follow the applicable government regulations and local guidelines
• Elecsys 2010, MODULAR ANALYTICS E170 or cobas e analyzer for quality control.
Accessories for Elecsys 2010 and cobas e 411 analyzers: Calculation
• REF 11662988122, ProCell, 6 x 380 mL system buffer The analyzer automatically calculates the analyte concentration of
• REF 11662970122, CleanCell, 6 x 380 mL measuring cell cleaning solution each sample (either in IU/mL or kIU/L).
• REF 11930346122, Elecsys SysWash, 1 x 500 mL washwater additive Limitations - interference
• REF 11933159001, Adapter for SysClean The assay is unaffected by icterus (bilirubin < 1129 µmol/L or < 66 mg/dL),
• REF 11706802001, Elecsys 2010 AssayCup, 60 x 60 reaction vessels hemolysis (Hb < 0.93 mmol/L or < 1.5 g/dL), lipemia (triglycerides
• REF 11706799001, Elecsys 2010 AssayTip, 30 x 120 pipette tips < 23.9 mmol/L or < 2100 mg/dL), and biotin < 40.9 nmol/L or < 10 ng/mL.
Accessories for MODULAR ANALYTICS E170, cobas e 601 Criterion: Recovery within ± 10 % of initial value.
and cobas e 602 analyzers: In patients receiving therapy with high biotin doses (i.e. > 5 mg/day), no sample
should be taken until at least 8 hours after the last biotin administration.
• REF 04880340190, ProCell M, 2 x 2 L system buffer
No interference was observed from rheumatoid factors up to
• REF 04880293190, CleanCell M, 2 x 2 L measuring cell cleaning solution a concentration of 1500 IU/mL.
• REF 03023141001, PC/CC-Cups, 12 cups to prewarm ProCell M In vitro tests were performed on 23 commonly used pharmaceuticals.
and CleanCell M before use No interference with the assay was found.
• REF 03005712190, ProbeWash M, 12 x 70 mL cleaning solution for In rare cases, interference due to extremely high titers of antibodies to
run finalization and rinsing during reagent change analyte-specific antibodies, streptavidin or ruthenium can occur. These
• REF 12102137001, AssayTip/AssayCup Combimagazine M, 48 magazines effects are minimized by suitable test design.
x 84 reaction vessels or pipette tips, waste bags For diagnostic purposes, the results should always be assessed in conjunction
• REF 03023150001, WasteLiner, waste bags with the patient’s medical history, clinical examination and other findings.
• REF 03027651001, SysClean Adapter M Limits and ranges
Accessories for all analyzers: Measuring range
• REF 11298500316, Elecsys SysClean, 5 x 100 mL system cleaning solution 5.00-600 IU/mL (defined by the lower detection limit and the maximum of the
master curve). Values below the detection limit are reported as < 5.00 IU/mL.
• REF 11298500160, Elecsys SysClean, 5 x 100 mL system
Values above the measuring range are reported as > 600 IU/mL.
cleaning solution (for USA)
Lower limits of measurement
Assay Lower detection limit
For optimum performance of the assay follow the directions given in < 5.00 IU/mL
this document for the analyzer concerned. Refer to the appropriate The detection limit represents the lowest analyte level that can be
operator’s manual for analyzer-specific assay instructions. distinguished from zero. It is calculated as the value lying two standard

Elecsys and cobas e analyzers 2/3 2010-10, V 3 English

05572339001V3

Anti-TPO
Antibodies to thyroid peroxidase
deviations above that of the lowest standard (master calibrator, References
standard 1 + 2 SD, repeatability study, n = 21). 1. Pfannenstiel P, Saller B. Schilddrüsenkrankheiten - Diagnose
und Therapie, 2nd edition. Berliner Medizinische Verlagsanstalt.
Dilution
1995;28-30,141,169-172,200-201.
Samples with anti-TPO concentrations above the measuring range can be
2. McIntosh RS, Asghar MS, Weetman AP. The antibody response in human
diluted manually with Elecsys Diluent Universal. The recommended dilution
autoimmune thyroid disease. Clin Sci 1997;(92)6:529-541.
is 1:5. The concentration of the diluted sample must be > 200 IU/mL.
3. Volpé R. Rational Use of Thyroid Function Tests.
After dilution, multiply the result by the dilution factor.
Crit Rev Clin Lab Sci 1997;34(5):405-438.
Please note: The autoantibodies are heterogeneous and this gives rise
4. Feldt-Rasmussen U. Analytical and clinical performance goals
to non-linear dilution phenomena for individual samples.
for testing autoantibodies to thyroperoxidase, thyroglobulin, and
Expected values thyrotropin receptor. Clin Chem 1996;42(1):160-163.
Extended studies with the Elecsys Anti-TPO assay performed on samples 5. Utiger RD. The pathogenesis of autoimmune thyroid disease.
from 208 healthy test subjects in 3 clinical centers in Austria and Germany N Eng J Med 1991;325:278-279.
showed a borderline value of 34 IU/mL for 95 % of the results. 6. Gutekunst R. Hashimoto-Thyreoiditis: Diagnostik und Verlaufskontrolle.
For detailed information about reference intervals in children, adolescents In: Börner W, Weinheimer B (Hrsg.): Schilddrüse 1989.
and pregnant women, refer to the brochure “Reference Intervals for Children Walter de Gruyter, Berlin, New York 1991;348-355.
and Adults”, REF English: 04640292, German: 04625889. 7. Occupational Safety and Health Standards: bloodborne pathogens.
This booklet also contains results of a detailed study about influencing factors (29 CFR Part 1910.1030). Fed. Register.
on thyroid parameters in a well characterized reference group of adults. 8. Council Directive (2000/54/EC). Official Journal of the European
Different inclusion and exclusion criteria were applied (e.g. sonographic results Communities No. L262 from Oct. 17, 2000.
(thyroid volume and density) as well as criteria according to the guidelines 9. Greiling H, Gressner AM. Lehrbuch der Klinischen Chemie und
of the National Academy of Clinical Biochemistry - NACB). Pathobiochemie. 3rd edition, Stuttgart; New York: Schattauer 1995:1012.
Each laboratory should investigate the transferability of the expected values to 10. Passing H, Bablok W, et al. A General Regression Procedure for Method
its own patient population and if necessary determine its own reference ranges. Transformation. J Clin Chem Clin Biochem 1988;26:783-790.

Specific performance data For further information, please refer to the appropriate operator’s manual
Representative performance data on the analyzers are given below. for the analyzer concerned, the respective application sheets, the product
Results obtained in individual laboratories may differ. information, and the package inserts of all necessary components.
FOR US CUSTOMERS ONLY: LIMITED WARRANTY
Precision
Roche Diagnostics warrants that this product will meet the specifications
Precision was determined using Elecsys reagents, pooled human sera, and
stated in the labeling when used in accordance with such labeling and
controls (repeatability: n = 21, intermediate precision: n = 21); total precision
will be free from defects in material and workmanship until the expiration
on MODULAR ANALYTICS E170 analyzer was determined in a modified
date printed on the label. THIS LIMITED WARRANTY IS IN LIEU OF ANY
protocol (EP5-A) of the CLSI (Clinical and Laboratory Standards Institute): 6
OTHER WARRANTY, EXPRESS OR IMPLIED, INCLUDING ANY IMPLIED
times daily for 10 days (n = 60). The following results were obtained:
WARRANTY OF MERCHANTABILITY OR FITNESS FOR PARTICULAR
Elecsys 2010 and cobas e 411 analyzers PURPOSE. IN NO EVENT SHALL ROCHE DIAGNOSTICS BE LIABLE FOR
Repeatabilityb Intermediate precision INCIDENTAL, INDIRECT, SPECIAL OR CONSEQUENTIAL DAMAGES.
Sample Mean SD CV Mean SD CV
IU/mL IU/mL % IU/mL IU/mL %
Human serum 1 15.3 1.07 7.0 12.4 3.02 24.4
COBAS, COBAS E, ELECSYS and MODULAR are trademarks of Roche.
Human serum 2 113 2.88 2.5 109 10.1 9.2 Other brand or product names are trademarks of their respective holders.
Human serum 3 269 11.4 4.2 308 21.9 7.1 Significant additions or changes are indicated by a change bar in the margin. Changes to reagent barcode
test parameters which have already been read in should be edited manually.
PC A-TPO1 25.9 1.45 5.6 25.4 3.35 13.2 © 2010, Roche Diagnostics
PC A-TPO2 112 3.78 3.4 116 8.83 7.6
b) Repeatability = within-run precision
Roche Diagnostics GmbH, Sandhofer Strasse 116, D-68305 Mannheim
MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 analyzers www.roche.com
Repeatability Intermediate precision Distribution in USA by:
Sample Mean SD CV Mean SD CV Roche Diagnostics, Indianapolis, IN
US Customer Technical Support 1-800-428-2336
IU/mL IU/mL % IU/mL IU/mL %
Human serum 1 21.3 1.34 6.3 20.8 1.97 9.5
Human serum 2 51.2 2.61 5.1 53.1 3.25 6.1
Human serum 3 473 12.7 2.7 455 19.1 4.2
PC A-TPO1 19.4 1.07 5.5 22.0 2.02 9.2
PC A-TPO2 100 2.57 2.6 103 4.39 4.3
Method comparison
A comparison of the Elecsys Anti-TPO assay (y) with a commercially available
anti-TPO test (x) using clinical samples gave the following correlations:
Number of samples measured: 50
Passing/Bablok10 Linear regression
y = 0.77x + 2.94 y = 0.63x + 17.1
τ = 0.785 r = 0.899
The sample concentrations were between approx. 12 and 460 IU/mL.
Analytical specificity
No influence with human autoantibodies to thyroglobulin
(< 394 IU/mL) was detectable.

2010-10, V 3 English 3/3 Elecsys and cobas e analyzers