ms_08849544190V1.

FXII
Factor XII

cobas t 511
08849544190 54 System‑ID 07 2003 6
cobas t 711

English Reagent handling

System information The reagent in the cassette has been assembled into a ready‑for‑use unit
(cobas t pack).
Short name ACN (application code number) All information required for correct operation is available via the cobas link.
FXII 28460 Storage and stability
Intended use Store at 2‑8 °C.
Human plasma immunodepleted of Factor XII for the quantitative Store the cobas t pack upright.
determination of Factor XII activity in citrated plasma, based on the The unopened cobas t pack is stable up to the stated expiration date.
activated partial thromboplastin time (aPTT) assay, on the indicated
cobas t analyzers. Stability of the opened cobas t pack:
Summary on the cobas t analyzer for each vial: 8 hours after
Factor XII (FXII) is circulating in plasma as a single chain zymogen. Upon reconstitution
contact with negatively charged surfaces, FXII is converted into its two-
chained active form (FXIIa). FXIIa initiates the intrinsic coagulation cascade Do not freeze.
via activation of FXI as well as the kallikrein-kinin system through Specimen collection and preparation
conversion of prekallikrein to kallikrein.1 Kallikrein reciprocally activates FXII Only the specimens listed below were tested and found acceptable:
in a positive feedback loop thus enhancing the intrinsic coagulation 3.2 % citrated human plasma.
response, which eventually leads to the formation of a fibrin clot. FXIIa also
counteracts the formation of stable fibrin clots by converting plasminogen to Use standard sampling tubes made of plastic or siliconized glass. Strictly
plasmin through activation of the urokinase-type plasminogen activator, observe the ratio of blood (9 parts) to sodium citrate solution 0.11 M
thus promoting fibrinolysis.1 Activation of FXII can also be triggered by non- (1 part).6,7
physiologic contact activators, like silicates or sulphated polysaccharides as The sample types listed were tested with a selection of sample collection
well as non-biological surfaces including metallic heart valves, catheters tubes that were commercially available at the time of testing, i.e. not all
and extracorporeal circuits like cardiopulmonary bypass and available tubes of all manufacturers were tested. Sample collection systems
hemodialysis.2,3 from various manufacturers may contain differing materials which could
Test principle affect the test results in some cases. When processing samples in primary
tubes (sample collection systems), follow the instructions of the tube
FXII activity is determined with an aPTT‑type assay. In order to make FXII manufacturer.
activity rate‑limiting, the patient plasma is diluted and added to a
FXII‑deficient human plasma, with a FXII level < 1 % of the norm, whereas Centrifuge 15 minutes at 2500 g or such that the platelet count is
all other coagulation factors are present at levels typically > 50 % of the < 10000 platelets/μL and assay samples within the given stability period.
norm. The resulting clotting time is interpreted by using a calibration curve,
obtained with dilutions of a calibrator plasma mixed with FXII‑deficient Stability:
human plasma. The extent of aPTT correction is proportional to the FXII at 15‑25 °C 4 hours
activity in the sample, which is determined quantitatively from the calibration
curve. at -20 °C (± 5 °C) 28 days
Reagents - working solutions Frozen plasma aliquots should be thawed within 5 minutes at 37 °C in a
waterbath, homogenized by carefully mixing without foam formation. Assay
cobas t pack thawed samples within 2 hours. Do not refreeze samples.
R2 Lyophilized FXII immunodepleted human plasma Materials provided
R2 is in position A, B and C. See “Reagents – working solutions” section.
Precautions and warnings Materials required (but not provided)
For in vitro diagnostic use for health care professionals. Exercise the ▪ 07575297190, Global Cal, 5 x 1 mL
normal precautions required for handling all laboratory reagents. ▪ 07539355190, Con N, 20 x 1 mL
Infectious or microbial waste: ▪ 07539665190, Con P, 20 x 1 mL
Warning: handle waste as potentially biohazardous material. Dispose of
waste according to accepted laboratory instructions and procedures. ▪ 07153678190, aPTT Lupus,600 T
Environmental hazards: ▪ 07154984190, CC 25mM, 50 mL
Apply all relevant local disposal regulations to determine the safe disposal. ▪ 07155042190, Owren B, 50 mL
Safety data sheet available for professional user on request.
▪ General laboratory equipment
All human material should be considered potentially infectious. All products
derived from human blood are prepared exclusively from the blood of ▪ Distilled or deionized water
donors tested individually and shown to be free from HBsAg and antibodies ▪ cobas t coagulation analyzer. See User Assistance of the analyzer
to HCV and HIV. The testing methods use assays that have been approved concerned for additionally required materials.
by the FDA or that are in compliance with the legal rules applicable to
placing in vitro diagnostic medical devices for human use on the market in Assay
the European Union. For optimum performance of the assay follow the directions given in this
However, as no testing method can rule out the potential risk of infection document. Refer to the appropriate User Assistance for analyzer‑specific
with absolute certainty, the material should be handled with the same level assay instructions.
of care as a patient specimen. In the event of exposure, the directives of the The performance of applications not validated by Roche is not warranted
responsible health authorities should be followed.4,5 and must be defined by the user.
Avoid foam formation in all reagents and sample types (specimens, Calibration
calibrators and controls). For calibration, use calibrator as listed in the "Materials required (but not
provided)" section.

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ms_08849544190V1.0

FXII
Factor XII

Calibration frequency: full calibration must be performed The Limit of Quantitation is defined as the lowest analyte concentration in a
sample that can be reproducibly measured with an intermediate precision
▪ once per lot of aPTT Lupus reagent or FXII-deficient plasma CV of ≤ 20 %.
▪ as required following quality control procedures.
Expected values
Traceability: This method has been standardized against the international
WHO/NIBSC FXII plasma standard. 52.7‑169 %
These values correspond to the 2.5th and 97.5th percentiles of results
Quality control obtained from a total of 199 human plasma samples.
Controls are required for checking the accuracy and reproducibility of the Each laboratory should investigate the transferability of the expected values
results. to its own patient population and if necessary determine its own reference
For quality control, use control kits as listed in the "Materials required (but ranges.
not provided)" section.
Specific performance data
The control intervals and limits should be adapted to each laboratory’s
individual requirements. Values obtained should fall within the defined Representative performance data on the analyzers are given below.
limits. Each laboratory should establish corrective measures to be taken if Results obtained in individual laboratories may differ.
values fall outside the defined limits. Precision
Follow the applicable government regulations and local guidelines for Repeatability and intermediate precision were determined using human
quality control. samples and controls in accordance with the CLSI (Clinical and Laboratory
Standards Institute) EP05 requirements (2 aliquots per run, 2 runs per day,
Calculation 21 days).14 The following results were obtained:
The cobas t analyzers automatically calculate the analyte activity of each
sample. Repeatability Intermediate
Conversion factor: 100 % = 1 IU/mL.8,9 precision
Limitations - interference Sample Mean SD CV SD CV
The effect of the following endogenous substances and pharmaceutical (%) (%) (%) (%) (%)
compounds on assay performance was tested. No impact on results was
observed up to the listed concentrations. Con N 98.4 1.83 1.9 2.43 2.5
Con P 38.7 0.783 2.0 1.03 2.7
Endogenous substances
Plasma 1 5.90 0.0981 1.7 0.147 2.5
Compound Concentration
Plasma 2 22.1 0.429 1.9 0.604 2.7
Conjugated bilirubin 5 mg/dL
Plasma 3 49.6 1.23 2.5 1.60 3.2
Unconjugated bilirubin 50 mg/dL
Plasma 4 71.1 1.36 1.9 2.10 3.0
Hemoglobin 500 mg/dL
Plasma 5 136 2.80 2.1 4.42 3.3
Intralipid 1200 mg/dL
Method comparison
Criterion: Recovery within ± 10.0 % of initial value.
A comparison of the FXII assay on the cobas t 711 analyzer (y) with an
The impact of lipemia, hemoglobin and bilirubin was tested according to automated coagulation assay (x) gave the following correlation:
Glick.10
Number of samples measured: 118
Drugs: No interference was found at therapeutic concentrations using
common drug panels except for heparin.11,12 Deming15
The fibrinolytic action of streptokinase alters the clotting time and thus the y = 0.984x + 1.43
FXII activity. The presence of fondaparinux and direct thrombin inhibitors
such as argatroban, bivalirudin and dabigatran, or factor Xa inhibitors, such r = 0.992
as edoxaban, rivaroxaban, and apixaban, in the sample influences the
assay results (reduced FXII activity) which can be of clinical importance. The FXII activities using the FXII reagent were between 5.69 and 145 %.
No significant interference was observed up to an Orbactiv concentration of References
30 mg/L. 1 Didiasova M, Wujak L, Schaefer L, Wygrecka M. Factor XII in
For diagnostic purposes, the results should always be assessed in coagulation, inflammation and beyond. Cell Signal. 2018
conjunction with the patient’s medical history, clinical examination and other Nov;51:257-265.
findings. 2 Naudin C, Burillo E, Blankenberg S, Butler L, Renné T. Factor XII
Extra wash cycle: The use of special wash steps is mandatory when Contact Activation. Semin Thromb Hemost. 2017 Nov;43(8):814-826.
certain test combinations are run together on cobas t analyzers. Refer to 3 Weitz JI, Fredenburgh JC. Factors XI and XII as Targets for New
the latest version of the carry over evasion list found with the CLEAN and Anticoagulants. Front Med (Lausanne). 2017 Feb 24;4:19.
Deproteinizer Method Sheets and the User Assistance for further
instructions. Where required, special wash/carry over evasion cycles must 4 Occupational Safety and Health Standards: Bloodborne pathogens.
be implemented prior to reporting results with this test. (29 CFR Part 1910.1030). Fed. Register.
Limits and ranges 5 Directive 2000/54/EC of the European Parliament and Council of
18 September 2000 on the protection of workers from risks related to
Measuring range exposure to biological agents at work.
1.00‑150 %
6 CLSI Document H21-A5, Vol.28, No.5, 2008. Collection, transport, and
For samples with concentrations > 150 to 250 % FXII activity, the rerun processing of blood specimens for testing plasma-based coagulation
function decreases the sample volume by a factor of 3 and the results are assays and molecular hemostasis assays; approved guideline, 5th
automatically multiplied by this factor. edition.
Lower limits of measurement 7 CLSI Document H3-A6. Procedures for the collection of diagnostic
Limit of Quantitation (LoQ) = 1.00 % blood specimens by venipuncture; approved standard - Sixth Edition,
The Limit of Quantitation was determined in accordance with the CLSI vol. 27, No. 26, 2007.
(Clinical and Laboratory Standards Institute) EP17‑A2 requirements.13

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Factor XII

8 Bristow AF, Barrowcliffe T, Bangham DR. Standardization of biological

medicines: the first hundred years, 1900-2000. Notes Rec R Soc Lond.
2006 Sep 22;60(3):271-89.
9 Hubbard AR. International biological standards for coagulation factors
and inhibitors. Semin Thromb Hemost. 2007 Apr;33(3):283-9.
10 Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of
Interferences in Clinical Chemistry Instrumentation.
Clin Chem 1986;32:470-475.
11 Breuer J. Report on the Symposium “Drug effects in Clinical Chemistry
Methods”. Eur J Clin Chem Clin Biochem 1996;34:385-386.
12 Sonntag O, Scholer A. Drug interference in clinical chemistry:
recommendation of drugs and their concentrations to be used in drug
interference studies. Ann Clin Biochem 2001;38:376-385.
13 CLSI Document EP17-A2. Evaluation of Detection Capability for
Clinical Laboratory Measurement Procedures. Vol. 32, No. 8, 2012.
Approved standard, 2nd Edition.
14 CLSI Document EP05-A3. Evaluation of Precision of Quantitative
Measurement Procedures. Vol. 24, No. 25, 2014. Approved guideline,
3rd Edition.
15 Martin RF. General Deming Regression for Estimating Systematic Bias
and its Confidence Interval in Method Comparison Studies. Clinical
Chemistry 2000;46(1):100-104.
A point (period/stop) is always used in this Method Sheet as the decimal
separator to mark the border between the integral and the fractional parts of
a decimal numeral. Separators for thousands are not used.
For further information, please refer to the appropriate User Assistance for
the relevant analyzer and Method Sheets of all necessary components.
Any serious incident that has occurred in relation to the device shall be
reported to the manufacturer and the competent authority of the Member
State in which the user and/or the patient is established.
Symbols
Roche Diagnostics uses the following symbols and signs in addition to
those listed in the ISO 15223‑1 standard (for USA: see dialog.roche.com for
definition of symbols used):
Contents of kit
Analyzers/Instruments on which reagents can be used
Reagent
Calibrator
Volume for reconstitution
GTIN Global Trade Item Number

COBAS and COBAS T are trademarks of Roche.

All other product names and trademarks are the property of their respective owners.
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