ms_11776223322V17.

CA 125 II
Cancer Antigen 125

Elecsys 2010
MODULAR ANALYTICS E170
11776223 322 100 cobas e 411
cobas e 601
cobas e 602

English malignancies of the endometrium, breast, gastrointestinal tract, and various

other malignancies.
Please note Although CA 125 is a relatively unspecific marker,8,9,10,11,12 it is today the
The measured CA 125 value of a patient’s sample can vary depending on most important tumor marker for monitoring the therapy and progress of
the testing procedure used. The laboratory finding must therefore always patients with serous ovarian carcinoma. At primary diagnosis the sensitivity
contain a statement on the CA 125 assay method used. CA 125 values of CA 125 depends on the FIGO stage (FIGO = Federation of Gynecology
determined on patient samples by different testing procedures cannot be and Obstetrics); higher tumor stages are associated with higher CA 125
directly compared with one another and could be the cause of erroneous levels.13
medical interpretations. If there is a change in the CA 125 assay The diagnostic sensitivity and specificity of the Elecsys CA 125 II test was
procedure used while monitoring therapy, then the CA 125 values calculated by comparing ovarian carcinoma patients at primary diagnosis
obtained upon changing over to the new procedure must be confirmed by (FIGO stage I to IV) with patients suffering from benign gynecological
parallel measurements with both methods. diseases. At a cutoff value of 65 U/mL, the sensitivity is 79 % (at a low
Intended use specificity of 82 %). The cutoff level has to be raised if higher specificity is
desired. The optimal clinical value is reached at 150 U/mL (sensitivity 69 %,
Immunoassay for the in vitro quantitative determination of OC 125 reactive specificity 93 %). If the specificity is 95 %, in accordance with the
determinants in human serum and plasma. recommendations of van Dalen, et al.,14 a sensitivity of 63 % is obtained
These determinants are associated with a high molecular weight (cutoff 190 U/mL).
glycoprotein in serum and plasma of women with primary epithelial invasive
ovarian cancer (excluding those with cancer of low malignant potential). Test principle
Sandwich principle. Total duration of assay: 18 minutes.
This assay is indicated for use as an aid in the detection of residual or
recurrent ovarian carcinoma in patients who have undergone first‑line ▪ 1st incubation: 20 µL of sample, a biotinylated monoclonal
therapy and would be considered for second-look procedures. This assay is CA 125‑specific antibody, and a monoclonal CA 125‑specific antibody
further indicated for serial measurement of CA 125 to aid in the labeled with a ruthenium complexa) form a sandwich complex.
management of cancer patients. ▪ 2nd incubation: After addition of streptavidin-coated microparticles, the
This assay is also intended to be used in conjunction with the Elecsys HE4 complex becomes bound to the solid phase via interaction of biotin and
assay as part of ROMA (Risk Of Ovarian Malignancy Algorithm) for the risk streptavidin.
assessment of ovarian cancer in premenopausal and postmenopausal
women presenting with pelvic mass. ▪ The reaction mixture is aspirated into the measuring cell where the
microparticles are magnetically captured onto the surface of the
The electrochemiluminescence immunoassay “ECLIA” is intended for use electrode. Unbound substances are then removed with
on Elecsys and cobas e immunoassay analyzers. ProCell/ProCell M. Application of a voltage to the electrode then induces
Summary chemiluminescent emission which is measured by a photomultiplier.
CA 125 belongs to the family of hybridoma‑defined tumor markers. The ▪ Results are determined via a calibration curve which is instrument-
values measured are defined by the use of the monoclonal antibody (MAb) specifically generated by 2‑point calibration and a master curve provided
OC 125. via the reagent barcode.
The antigenic determinant CA 125 is located on a high‑molecular weight a) Tris(2,2'-bipyridyl)ruthenium(II)-complex (Ru(bpy) )
glycoprotein (200‑1000 kD) isolated from cell culture or serum. The Reagents - working solutions
antigenic determinant CA 125 has a protein structure with associated The reagent rackpack is labeled as CA125 II.
carbohydrate side-chains.1
MAb OC 125 was obtained from lymphocytes from mice that had been M Streptavidin-coated microparticles (transparent cap), 1 bottle, 6.5 mL:
immunized with OVCA (ovarian carcinoma cell line) 433, an Streptavidin-coated microparticles 0.72 mg/mL; preservative.
adenocarcinoma cell line from the ovary.2 In the Elecsys test, OC 125 is
used as a detection antibody. MAb M 11 is used as the capture antibody R1 Anti-CA 125-Ab~biotin (gray cap), 1 bottle, 9 mL:
(solid-phase antibody); this has been employed in second-generation Biotinylated monoclonal anti‑CA 125 antibody (M 11; mouse) 1 mg/L;
CA 125 assays since 1992. phosphate buffer 100 mmol/L, pH 7.4; preservative.
CA 125 is found in a high percentage of non-mucinous ovarian tumors of
epithelial origin3 and can be detected in serum.4,5 It does not occur on the R2 Anti-CA 125-Ab~Ru(bpy) (black cap), 1 bottle, 9 mL:
surface epithelium of normal ovaries (adult and fetal). Ovarian carcinoma Monoclonal anti‑CA 125 antibody (OC 125; mouse) labeled with
accounts for about 20 % of gynecological tumors; the incidence is ruthenium complex 1 mg/L; phosphate buffer 100 mmol/L, pH 7.4;
15/100000.6 preservative.
CA 125 has been found in the amniotic fluid and in the coelomic epithelium;
both of these tissues are of fetal origin. In tissues of adult origin, the Precautions and warnings
presence of CA 125 has been demonstrated in the epithelium of the For in vitro diagnostic use.
oviduct, in the endometrium and in the endocervix.7 Exercise the normal precautions required for handling all laboratory
Elevated values are sometimes found in various benign gynecological reagents.
diseases such as ovarian cysts, ovarian metaplasia, endometriosis, uterus Disposal of all waste material should be in accordance with local guidelines.
myomatosus or cervicitis. Slight elevations of this marker may also occur in Safety data sheet available for professional user on request.
early pregnancy and in various benign diseases (e.g. acute and chronic Avoid foam formation in all reagents and sample types (specimens,
pancreatitis, benign gastrointestinal diseases, renal insufficiency, calibrators and controls).
autoimmune diseases and others). Markedly elevated levels have been Reagent handling
found in benign liver diseases such as cirrhosis and hepatitis. Extreme
elevations can occur in any kind of ascites due to malignant and benign The reagents in the kit have been assembled into a ready‑for‑use unit that
diseases. Although the highest CA 125 values occur in patients suffering cannot be separated.
from ovarian carcinoma, clearly elevated values are also observed in

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CA 125 II
Cancer Antigen 125

All information required for correct operation is read in from the respective ▪  11706799001, Elecsys 2010 AssayTip, 30 x 120 pipette tips
reagent barcodes. Accessories for MODULAR ANALYTICS E170, cobas e 601 and
Storage and stability cobas e 602 analyzers:
Store at 2‑8 °C. ▪  04880340190, ProCell M, 2 x 2 L system buffer
Do not freeze. ▪  04880293190, CleanCell M, 2 x 2 L measuring cell cleaning
Store the Elecsys reagent kit upright in order to ensure complete solution
availability of the microparticles during automatic mixing prior to use. ▪  03023141001, PC/CC‑Cups, 12 cups to prewarm ProCell M and
CleanCell M before use
Stability:
▪  03005712190, ProbeWash M, 12 x 70 mL cleaning solution for run
unopened at 2‑8 °C up to the stated expiration date finalization and rinsing during reagent change
after opening at 2‑8 °C 12 weeks ▪  12102137001, AssayTip/AssayCup Combimagazine M,
on the analyzers 6 weeks 48 magazines x 84 reaction vessels or pipette tips, waste bags
▪  03023150001, WasteLiner, waste bags
Specimen collection and preparation
▪  03027651001, SysClean Adapter M
Only the specimens listed below were tested and found acceptable.
Accessories for all analyzers:
Serum collected using standard sampling tubes or tubes containing
separating gel. ▪  11298500316, Elecsys SysClean, 5 x 100 mL system cleaning
solution
Li-, Na-, NH ‑heparin, K2‑EDTA, K3‑EDTA and sodium citrate plasma as
well as plasma tubes containing separating gel. When sodium citrate Assay
plasma is used, the results must be corrected by + 10 %. For optimum performance of the assay follow the directions given in this
Criterion: Recovery within 90‑110 % of serum value or slope document for the analyzer concerned. Refer to the appropriate operator’s
0.9‑1.1 + intercept within < ± 2x analytical sensitivity (LDL) + coefficient of manual for analyzer‑specific assay instructions.
correlation > 0.95. Resuspension of the microparticles takes place automatically prior to use.
Stable for 5 days at 2‑8 °C, 3 months at ‑20 °C.15 Read in the test‑specific parameters via the reagent barcode. If in
exceptional cases the barcode cannot be read, enter the 15‑digit sequence
The sample types listed were tested with a selection of sample collection of numbers.
tubes that were commercially available at the time of testing, i.e. not all
available tubes of all manufacturers were tested. Sample collection systems Bring the cooled reagents to approximately 20 °C and place on the reagent
from various manufacturers may contain differing materials which could disk (20 °C) of the analyzer. Avoid foam formation. The system
affect the test results in some cases. When processing samples in primary automatically regulates the temperature of the reagents and the
tubes (sample collection systems), follow the instructions of the tube opening/closing of the bottles.
manufacturer. Calibration
Centrifuge samples containing precipitates before performing the assay. Traceability: This method has been standardized against the Enzymun‑Test
Do not use heat‑inactivated samples. CA 125 II method. This in turn has been standardized against the CA 125 II
Do not use samples and controls stabilized with azide. RIA from Fujirebio Diagnostics.
Ensure the samples, calibrators and controls are at 20‑25 °C prior to Every Elecsys reagent set has a barcoded label containing specific
measurement. information for calibration of the particular reagent lot. The predefined
master curve is adapted to the analyzer using the relevant CalSet.
Due to possible evaporation effects, samples, calibrators and controls on
the analyzers should be analyzed/measured within 2 hours. Calibration frequency: Calibration must be performed once per reagent lot
using fresh reagent (i.e. not more than 24 hours since the reagent kit was
Materials provided registered on the analyzer). Renewed calibration is recommended as
See “Reagents – working solutions” section for reagents. follows:
Materials required (but not provided) ▪ after 8 weeks when using the same reagent lot
▪  11776240322, CA 125 II CalSet, 4 x 1 mL ▪ after 7 days (when using the same reagent kit on the analyzer)
▪  11776452122, PreciControl Tumor Marker, for 2 x 3 mL each of ▪ as required: e.g. quality control findings outside the defined limits
PreciControl Tumor Marker 1 and 2 Quality control
▪  11732277122, Diluent Universal, 2 x 16 mL sample diluent or For quality control, use PreciControl Tumor Marker.
 03183971122, Diluent Universal, 2 x 36 mL sample diluent In addition, other suitable control material can be used.
▪ General laboratory equipment Controls for the various concentration ranges should be run individually at
▪ Elecsys 2010, MODULAR ANALYTICS E170 or cobas e analyzer least once every 24 hours when the test is in use, once per reagent kit, and
For epithelial ovarian cancer risk assessment with ROMA (Risk of Ovarian following each calibration.
Malignancy Algorithm): The control intervals and limits should be adapted to each laboratory’s
individual requirements. Values obtained should fall within the defined
▪  05950929190, HE4, 100 tests limits. Each laboratory should establish corrective measures to be taken if
▪  05950945190, HE4 CalSet, for 4 x 1 mL values fall outside the defined limits.
▪  05950953190, PreciControl HE4, for 2 x 1 mL each of PreciControl Follow the applicable government regulations and local guidelines for
HE4 1 and 2 quality control.
▪  03609987190, Diluent MultiAssay, 2 x 16 mL sample diluent Calculation
Accessories for Elecsys 2010 and cobas e 411 analyzers: The analyzer automatically calculates the analyte concentration of each
sample (either in U/mL, U/L or kU/L).
▪  11662988122, ProCell, 6 x 380 mL system buffer
Limitations - interference
▪  11662970122, CleanCell, 6 x 380 mL measuring cell cleaning
solution The assay is unaffected by icterus (bilirubin < 1129 μmol/L or < 66 mg/dL),
hemolysis (Hb < 2.0 mmol/L or < 3.2 g/dL), lipemia (Intralipid < 2000 mg/dL)
▪  11930346122, Elecsys SysWash, 1 x 500 mL washwater additive and biotin (< 143 nmol/L or < 35 ng/mL).
▪  11933159001, Adapter for SysClean Criterion: Recovery within ± 10 % of initial value.
▪  11706802001, Elecsys 2010 AssayCup, 60 x 60 reaction vessels
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CA 125 II
Cancer Antigen 125

Samples should not be taken from patients receiving therapy with high Elecsys 2010 and cobas e 411 analyzers
biotin doses (i.e. > 5 mg/day) until at least 8 hours following the last biotin
administration. Repeatability Intermediate
precision
No interference was observed from rheumatoid factors up to a
concentration of 1200 IU/mL. Sample Mean SD CV SD CV
There is no high‑dose hook effect at CA 125 concentrations up to U/mL U/mL % U/mL %
50000 U/mL. Human serum 1 7.83 0.26 3.3 0.33 4.2
In vitro tests were performed on 27 commonly used pharmaceuticals. No
interference with the assay was found. Human serum 2 38.3 0.82 2.1 1.17 3.1
In rare cases, interference due to extremely high titers of antibodies to Human serum 3 70.8 1.46 2.1 1.75 2.5
analyte‑specific antibodies, streptavidin or ruthenium can occur. These PreciControl TMb)1 39.0 0.75 1.9 0.99 2.5
effects are minimized by suitable test design.
For diagnostic purposes, the results should always be assessed in PreciControl TM2 121.4 1.71 1.4 3.28 2.7
conjunction with the patient’s medical history, clinical examination and other b) TM = Tumor Marker
findings.
MODULAR ANALYTICS E170, cobas e 601 and cobas e 602 analyzers
Limits and ranges
Measuring range Repeatability Intermediate precision
0.600‑5000 U/mL (defined by the lower detection limit and the maximum of Sample Mean SD CV Mean SD CV
the master curve). Values below the lower detection limit are reported as U/mL U/mL % U/mL U/mL %
< 0.600 U/mL. Values above the measuring range are reported as
> 5000 U/mL (or up to 25000 U/mL for 5‑fold diluted samples). Human serum 1 21.1 0.23 1.1 20.1 0.37 1.8
Lower limits of measurement Human serum 2 198 1.39 0.7 199 3.43 1.7
Lower detection limit of the test Human serum 3 1816 29.3 1.6 1786 45.4 2.5
Lower detection limit: 0.600 U/mL PreciControl TM1 51.1 0.48 0.9 50.1 0.78 1.6
The lower detection limit represents the lowest measurable analyte level
that can be distinguished from zero. It is calculated as the value lying two PreciControl TM2 115 1.23 1.1 116 1.68 1.5
standard deviations above that of the lowest standard (master calibrator,
standard 1 + 2 SD, repeatability study, n = 21). Method comparison
A comparison of the Elecsys CA 125 II assay (y) with Fujirebio
Dilution Diagnostics CA 125 II RIA (x) using clinical samples gave the following
Samples with CA 125 concentrations above the measuring range can be correlations.
diluted with Diluent Universal. The recommended dilution is 1:5 (either Number of samples measured: 139
automatically by the MODULAR ANALYTICS E170, Elecsys 2010 or
cobas e analyzers or manually). The concentration of the diluted sample Passing/Bablok16 Linear regression
must be > 1000 U/mL.
y = 0.93x + 5.57 y = 0.96x + 5.82
After manual dilution, multiply the result by the dilution factor.
After dilution by the analyzers, the MODULAR ANALYTICS E170, τ = 0.81 r = 0.981
Elecsys 2010 and cobas e software automatically takes the dilution into The sample concentrations were between approximately 4 and 500 U/mL.
account when calculating the sample concentration.
Analytical specificity
Note: In rare cases, sample-dependent non-linearity upon dilution is seen
with samples having analyte levels beyond the measuring range. The Elecsys CA 125 II tumor marker assay is based on the monoclonal
M 11 and OC 125 antibodies which are only available from Fujirebio
Expected values Diagnostics, its licensees and its representatives. The performance
Studies using the Elecsys CA 125 II assay in 593 samples from healthy characteristics of test procedures using these antibodies cannot be
females (pre- and postmenopausal) yielded a value of 35 U/mL assumed for test methods using other antibodies.
(95th percentile). Values > 35 U/mL indicate an increased probability for References
residual or recurrent ovarian carcinoma in patients treated for primary
epithelial invasive ovarian cancer. 1 Davis HM, Zurawski VR Jr, Bast RC Jr, et al. Characterization of the
CA 125 antigen associated with human epithelial ovarian carcinomas.
Each laboratory should investigate the transferability of the expected values Cancer Research 1986;46:6143-6148.
to its own patient population and if necessary determine its own reference
ranges. 2 Bast RC, Feeney M, Lazarus H, et al. Reactivity of a monoclonal
antibody with human ovarian carcinoma. J Clin Invest 1981;68:1331.
Risk estimation in patients with pelvic mass
3 Kabawat SE, Bast RC Jr, Knapp RC, et al. Immunopathologic
For risk estimation with ROMA see package insert of the Elecsys HE4 characterization of a monoclonal antibody that recognizes common
assay. surface antigens of human ovarian tumors of serous, endometroid and
Specific performance data clear cell types. Am J Clin Pathol 1983;79:98-104.
Representative performance data on the analyzers are given below. 4 Bast RC, Klug TL, St. John E, et al. A radioimmunoassay using a
Results obtained in individual laboratories may differ. monoclonal antibody to monitor the course of epithelial ovarian cancer.
Precision N Engl J Med 1983;309:883-887.
Precision was determined using Elecsys reagents, pooled human sera and 5 Klug TL, Bast RC Jr, Niloff JM, et al. Monoclonal antibody
controls in a modified protocol (EP5‑A) of the CLSI (Clinical and Laboratory immunoradiometric assay for an antigenic determinant (CA 125)
Standards Institute): 6 times daily for 10 days (n = 60); repeatability on associated with human epithelial ovarian carcinomas. Cancer Res
MODULAR ANALYTICS E170 analyzer, n = 21. The following results were 1984;44:1048-1053.
obtained: 6 Hasholzner U, Baumgartner L, Stieber P, et al. Significance of the
tumor markers CA 125 II, CA 72-4, CASA and CYFRA 21-1 in ovarian
carcinoma. Anticancer Res 1994;14(6B):2743-2746.
7 Kabawat SE, Bast RC Jr, Bhan AK, et al. Tissue distribution of a
coelomic epithelium related antigen recognized by the monoclonal
antibody OC 125. Int J Gyn Path 1983;2:275-285.

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CA 125 II
Cancer Antigen 125

8 Daoud E, Bodor G, Weaver Ch, et al. (Washington University Case

Conference) CA-125 Concentrations in Malignant and Nonmalignant
Disease. Clin Chem 1991;37(11):1968-1974.
9 Kenemans P, Bon GG, Kessler A, et al. Multicenter Technical and
Clinical Evaluation of a Fully Automated Enzyme Immunoassay for CA
125. Clin Chem 1992;38/8:1466-1471.
10 Hasholzner U, Stieber P, Baumgartner L, et al. Methodological and
Clinical Evaluation of Three Automatized CA 125 Assays Compared
with CA 125 II RIA (Fujirebio). Tumordiagn Ther 1994;15:114-117.
11 Ruibal A, Encabo G, Martinez-Miralles E, et al. CA 125 seric levels in
nonmalignant pathologies. Bull Cancer (Paris) 1984;71(2):145-146.
12 Zahner J, Schmitz FJ, Schmitz G, et al. CA 125 - ein Tumormarker in
der Inneren Medizin? Lab med 1995;19:185-188.
13 Stieber P, Fateh-Moghadam A. Sensible Use of Tumormarkers. Dtsch:
Tumormarker und ihr sinnvoller Einsatz. ISBN 3-926725-07-9
dtsch/engl. Juergen Hartmann Verlag Marloffstein-Rathsberg (1993).
14 Van Dalen A. Quality Control and Standardization of Tumor Marker
Tests. Letter to the Editor: Tumor Biol 1993;14:131-135.
15 Guder WG, Narayanan S, Wisser H, et al. List of Analytes;
Preanalytical Variables. Brochure in: Samples: From the Patient to the
Laboratory. GIT-Verlag, Darmstadt 1996:8. ISBN 3-928865-22-6.
16 Bablok W, Passing H, Bender R, et al. A general regression procedure
for method transformation. Application of linear regression procedures
for method comparison studies in clinical chemistry, Part III.
J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.
For further information, please refer to the appropriate operator’s manual for
the analyzer concerned, the respective application sheets, the product
information and the Method Sheets of all necessary components (if
available in your country).
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.
CA 125 is a registered trademark of
Fujirebio Diagnostics, Inc.

Symbols
Roche Diagnostics uses the following symbols and signs in addition to
those listed in the ISO 15223‑1 standard.
Contents of kit
Analyzers/Instruments on which reagents can be used
Reagent
Calibrator
Volume after reconstitution or mixing

COBAS, COBAS E, ELECSYS, MODULAR and PRECICONTROL are trademarks of Roche. INTRALIPID is a
trademark of Fresenius Kabi AB.
All other product names and trademarks are the property of their respective owners.
Significant additions or changes are indicated by a change bar in the margin.
© 2014, Roche Diagnostics

Roche Diagnostics GmbH, Sandhofer Strasse 116, D-68305 Mannheim

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