30 426 08587 H - en - 2015/01

VIDAS® CA 125 II™ (125)

VIDAS CA 125 II is an automated quantitative test for use on the VIDAS family instruments, for the measurement of OC
125 antigenic determinants in human serum or plasma (lithium heparin or EDTA) using the ELFA technique (Enzyme
Linked Fluorescent Assay).

SUMMARY AND EXPLANATION PRINCIPLE

Tumor-associated OC 125 reactive antigenic The assay principle combines a 2-step enzyme
determinants are carried by high molecular weight immunoassay sandwich method with a final fluorescent
glycoproteins. The serum tumor marker assay, VIDAS CA detection (ELFA).
®
125 II, is a second generation test that uses monoclonal The Solid Phase Receptacle (SPR ) serves as the solid
OC 125 antibodies and a second monoclonal antibody, phase as well as the pipetting device for the assay.
M11, which can recognize another OC 125 antigenic Reagents for the assay are ready-to-use and pre-
determinant epitope [7, 8, 5, 2]. dispensed in the sealed reagent strips.
An increase in the assay value of CA 125 II is frequently All of the assay steps are performed automatically by the
found in certain types of cancer (ovary, breast…), but also instrument. The reaction medium is cycled in and out of
in certain non-cancerous pathologies [3, 10, 11, 6, 4]. the SPR several times.
Certain publications also describe the OC 125 antigenic The sample is cycled in and out of the SPR several times.
determinant as a marker for monitoring This operation enables the M11 antibody fixed onto the
endometriosis [1, 9]. interior wall of the SPR to capture the reactive antigenic
The CA 125 II assay level can decrease after therapy and determinants present in the sample. Unbound
increase in cases of relapse, residual disease and components are eliminated during the washing steps.
metastasis. The VIDAS CA 125 II assay is used as an Alkaline phosphatase-labeled OC 125 antibody
additional test for the prognosis and therapeutic (conjugate) is then incubated in the SPR where it binds
monitoring of patients with diagnosed malignant tumors. A with the OC 125 reactive antigenic determinants.
decrease in the CA 125 II assay level can indicate a Unbound conjugate is then eliminated during the washing
positive response to therapy and therefore good steps.
prognosis. A constant increase in the CA 125 II assay During the final detection step, the substrate (4-Methyl-
value often reflects evolution of the tumor and a poor umbelliferyl phosphate) is cycled in and out of the SPR.
response to therapy. The conjugate enzyme catalyzes the hydrolysis of this
substrate into a fluorescent product (4-Methyl-
umbelliferone) the fluorescence of which is measured at
450 nm. The intensity of the fluorescence is proportional
to the concentration of OC 125 reactive antigenic
determinants present in the sample.
At the end of the assay, results are automatically
calculated by the instrument in relation to the calibration
curve stored in memory, and then printed out.

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VIDAS CA 125 II™ (125) 08587 H - en - 2015/01

CONTENT OF THE KIT (30 TESTS) – RECONSTITUTION OF REAGENTS:

30 CA 125 II strips STR Ready-to-use.
30 CA 125 II SPRs SPR Ready-to-use.
Interior of SPRs coated with a monoclonal M 11 antibody (mouse).
CA 125 II control C1 Reconstitute using 2 ml of distilled water. Wait for 5 to 10 minutes before mixing.
1 x 2 ml (lyophilized) After reconstitution, stable for 48 hours at 2-8°C or 7 months at - 25 + 6°C. 5
freeze/thaw cycles are possible.
Human serum* + OC 125 antigenic determinants (human origin) + chemical
stabilizers.
MLE data indicate the confidence interval in U/mL ("Control C1 Dose Value
Range").
CA 125 II calibrator S1 Reconstitute using 3 ml of distilled water. Wait for 5 to 10 minutes before mixing.
1 x 3 ml (lyophilized) After reconstitution, stable for 48 hours at 2-8°C or 7 months at - 25 + 6°C. 5
freeze/thaw cycles are possible.
Human serum* + OC 125 antigenic determinants (human origin) + chemical
stabilizers.
MLE data indicate the concentration in U/mL (“Calibrator (S1) Dose Value”) and the
confidence interval in “Relative Fluorescence Value” (“Calibrator (S1) RFV Range”).
CA 125 II diluent R1 Ready-to-use.
1 x 5 ml (liquid) Bovine albumin + 0.9 g/l sodium azide.
Specifications for the factory master data required to calibrate the test:
• MLE data (Master Lot Entry) provided in the kit,
or
• MLE bar code printed on the box label.
1 Package insert provided in the kit or downloadable from www.biomerieux.com/techlib.
* This product has been tested and shown to be negative for HBs antigen, and antibodies to HIV1, HIV2 and HCV. However, since no
existing test method can totally guarantee their absence, this product must be treated as potentially infectious. Therefore, usual safety
procedures should be observed when handling.

The SPR
The strip
The interior of the SPR is coated during production with
mouse monoclonal M11 antibody. Each SPR is identified The strip consists of 10 wells covered with a labeled, foil
by the "125" code. Only remove the required number of seal. The label comprises a bar code which mainly
SPRs from the pouch and carefully reseal the pouch indicates the assay code, kit lot number and expiration
after opening. date. The foil of the first well is perforated to facilitate the
introduction of the sample. The last well of each strip is a
cuvette in which the fluorometric reading is performed.
The wells in the center section of the strip contain the
various reagents required for the assay.
Description of the CA 125 II strip:
Wells Reagents
1 Sample well.
2-3-4 Empty wells.
5 Conjugate: Alkaline phosphatase-labeled OC 125 antibody + 0.9 g/l sodium azide (400 µl).
6-7 Wash buffer: Tris (0.01 mol/l, pH 7.4) + Tween (0.05%) + NaCl (0.4 mol/l) + 0.9 g/l sodium azide (600 µl).
8 Diluent: Potassium phosphate (0.045 mol/l) + calf serum (10%) + 0.9 g/l sodium azide (400 µl).
9 Wash buffer: Diethanolamine* (DEA) (1.1 mol/l or 11.5%) pH 9.8 + 1 g/l sodium azide (600 µl).
10 Cuvette with substrate: 4-Methyl-umbelliferyl phosphate (0.6 mmol/l)
+ diethanolamine** (0.62 mol/l or 6.6%, pH 9.2) + 1 g/l sodium azide (300 µl).

* Signal Word: DANGER

Hazard statement
H318 : Causes serious eye damage.
H373 : May cause damage to organs through prolonged or repeated exposure.
H315 : Causes skin irritation.
H302 : Harmful if swallowed.

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VIDAS CA 125 II™ (125) 08587 H - en - 2015/01

Precautionary statement
P280 :Wear protective gloves/protective clothing/eye protection/face protection.
P305 + P351 + P338: IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present
and easy to do. Continue rinsing.
P309 + P311 : IF exposed or if you feel unwell: Call a POISON CENTER or doctor/physician.
** Signal Word: DANGER

Hazard statement
H318 : Causes serious eye damage.
Precautionary statement
P280 :Wear protective gloves/protective clothing/eye protection/face protection.
P305 + P351 + P338: IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present
and easy to do. Continue rinsing.
For further information, refer to the Material Safety Data Sheet.
MATERIALS AND DISPOSABLES REQUIRED BUT NOT PROVIDED
- Pipette with disposable tip to dispense 2 ml, 3 ml and 200 µl.
- Powderless, disposable gloves.
- For other specific materials and disposables, please refer to the Instrument User’s Manual.
- Instrument of the VIDAS family.

WARNINGS AND PRECAUTIONS • Spills should be wiped up thoroughly after treatment

• For in vitro diagnostic use only. with liquid detergent or a solution of household bleach
• For professional use only. containing at least 0.5% sodium hypochlorite. See the
• This kit contains products of human origin. No User’s Manual for cleaning spills on or in the instrument.
known analysis method can totally guarantee the Do not autoclave solutions containing bleach.
absence of transmissible pathogenic agents. It is • The instrument should be regularly cleaned and
therefore recommended that these products be decontaminated (see the User’s Manual).
treated as potentially infectious and handled
observing the usual safety precautions (see STORAGE CONDITIONS
Laboratory Bosafety Manual - WHO - Geneva - latest • Store the VIDAS CA 125 II kit at 2-8°C.
Edition). • Do not freeze the strips, SPRs or diluent.
• This kit contains products of animal origin. Certified • Store all unused reagents at 2-8°C.
knowledge of the origin and/or sanitary state of the • After opening the kit, check that the SPR pouch is
animals does not totally guarantee the absence of correctly sealed and undamaged. If not, do not use the
transmissible pathogenic agents. It is therefore SPRs.
recommended that these products be treated as • Carefully reseal the pouch with the desiccant inside
potentially infectious and handled observing the usual after use to maintain stability of the SPRs and return
safety precautions (do not ingest or inhale). the complete kit to 2-8°C.
• Do not use the SPRs if the pouch is pierced. • If stored according to the recommended conditions, all
• Do not use visibly deteriorated STRs (damaged foil or components are stable until the expiration date indicated
plastic). on the label. Refer to the kit composition table for
• Do not use reagents after the expiration date indicated special storage conditions.
on the label.
• Do not mix reagents (or disposables) from different lots. SPECIMENS
• Use powderless gloves, as powder has been reported
to cause false results for certain enzyme immunoassay Specimen type and collection:
tests. Human serum or plasma (lithium heparin or EDTA).
• Kit reagents contain sodium azide which can react with It is recommended that each laboratory checks the
lead or copper plumbing to form explosive metal azides. compatibility of collection tubes used.
If any liquid containing sodium azide is disposed of in Samples containing impurities must be centrifuged before
the plumbing system, drains should be flushed with analysis.
water to avoid build-up. None of the following factors have been found to
• The wash buffer in well 9 contains a harmful agent significantly influence this assay:
(11.5% diethanolamine). Refer to the hazard statements - hemolysis (after spiking samples with hemoglobin, 0 to
"H" and the precautionary statements "P" above. 300 µmol/l (monomer)),
• The substrate in well 10 contains an irritant agent (6.6% - lipemia (after spiking samples with lipids, 0 to 10 mg/ml
diethanolamine). Refer to the hazard statements "H" equivalent in triglycerides),
and the precautionary statements "P" above. - bilirubinemia (after spiking samples with bilirubin, 0 to
364 µmol/l).

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However, it is recommended not to use samples that are 9. The assay will be completed within approximately 60
clearly hemolyzed, lipemic or icteric and, if possible, to minutes. After the assay is completed, remove the
collect a new sample. SPRs and strips from the instrument.
10. Dispose of the used SPRs and strips into an
Specimen stability:
appropriate recipient.
Samples can be stored at 2-8°C in stoppered tubes for up
to 48 hours, or freeze the sera or plasma for 12 months at RESULTS AND INTERPRETATION
- 25 + 6°C. Avoid successive freezing and thawing. Once the assay is completed, results are analyzed
automatically by the computer. Fluorescence is measured
INSTRUCTIONS FOR USE
twice in the Reagent Strip’s reading cuvette for each
For complete instructions, see the User’s Manual. sample tested. The first reading is a background reading
Reading Master lot data of the substrate cuvette before the SPR is introduced into
the substrate. The second reading is taken after
Before each new lot of reagents is used, enter the incubating the substrate with the enzyme remaining on
specifications (or factory master data) into the instrument the interior of the SPR. The RFV (Relative Fluorescence
using the master lot entry (MLE) data. Value) is calculated by subtracting the background
If this operation is not performed before initiating the reading from the final result. This calculation appears on
tests, the instrument will not be able to print results. the result sheet. The results are automatically calculated
by the instrument using calibration curves which are
Note: the master lot data need only be entered once
stored by the instrument (4-parameter logistic model) and
for each lot.
the concentrations are expressed in U/mI.
It is possible to enter MLE data manually or
automatically depending on the instrument (refer to the Samples with a CA 125 II test value concentration greater
User’s Manual). than 600 U/ml should be reassayed after being diluted as
weakly as possible with the CA 125 II diluent (R1). If the
Calibration dilution factor has not been entered when the Work List
Calibration, using the calibrator provided in the kit, must was created (see User’s Manual), multiply the result by
be performed each time a new lot of reagents is opened, the dilution factor to obtain the sample concentration.
after the master lot data have been entered. Calibration Interpretation of test results should be made taking into
should then be performed every 14 days. This operation consideration the patient’s history, and the results of any
provides instrument-specific calibration curves and other tests performed.
compensates for possible minor variations in assay signal
throughout the shelf-life of the kit. QUALITY CONTROL
The calibrator, identified by S1, must be tested in A control is included in each VIDAS CA 125 II kit. This
duplicate (see User’s Manual). The calibrator value must control must be performed immediately after opening a
be within the set RFV "Relative Fluorescence Value" new kit to ensure that reagent performance has not been
range. If this is not the case, recalibrate. altered. Each calibration must also be checked using
Procedure these controls. The instrument will only be able to check
the control value if it is identified by C1.
1. Only remove the required reagents from the Results cannot be validated if the control value deviates
refrigerator and allow them to come to room from the expected values.
temperature for at least 30 minutes.
2. Use one "125" strip and one "125" SPR for each Note
sample, control or calibrator to be tested. Make sure It is the responsibility of the user to perform Quality
the storage pouch has been carefully resealed Control in accordance with any local applicable
after the required SPRs have been removed. regulations.
3. The test is identified by the "125" code on the LIMITATIONS OF THE METHOD
instrument. The calibrator must be identified by "S1",
and tested in duplicate. If the control is to be tested, it Interference may be encountered with certain sera
should be identified by "C1". containing antibodies directed against reagent
components. For this reason, assay results should be
4. Mix the calibrator and/or the control and samples
interpreted taking into consideration the patient's history,
using a vortex-type mixer (for serum or plasma
and the results of any other tests performed.
separated from the pellet).
The serum tumor marker assay, VIDAS CA 125 II, is
5. For this test, the calibrator, control, and sample based on the use of the monoclonal antibodies OC 125
test portion is 200 µl. and M11 supplied exclusively by Fujirebio Diagnostics,
6. Insert the "125" SPRs and "125" strips into the Inc., their distributors and licensed organizations. Methods
instrument. Check to make sure the color labels with using antibodies other than OC 125 and M11 may give
the assay code on the SPRs and the Reagent Strips different clinical results.
match. The VIDAS CA 125 II assay cannot be used as a
7. Initiate the assay as directed in the User’s Manual. All screening test for cancer.
the assay steps are performed automatically by the It is advised not to perform CA 125 II assays in patients
instrument. who have received a contrast agent in the previous 24
8. Reclose the vials and return them to the required hours [12].
temperature after pipetting.

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RANGE OF EXPECTED VALUES

These figures are given as a guide; it is recommended that each laboratory establish its own reference values from a
rigorously selected population.
The expected values were determined using 1181 samples collected from 451 healthy subjects, 520 subjects with a
diagnosed malignant pathology, and 211 subjects with a diagnosed non-malignant pathology.
EXPECTED No. Zone of values in U/ml

VALUES of 0-35 35-50 50-250 250-600 > 600

subjects % N % N % N % N % N
HEALTHY 451 99.1% 447 0.2% 1 0.7% 3 0.0% 0 0.0% 0
SUBJECTS
Malignant
pathologies
Ovarian 252 36.9 93 4.8 12 19.8 50 9.5 24 29.0 73
Ovarian at initial
stage 25 12.0 3 0.0 0 16.0 4 20.0 5 52.0 13
Breast 75 88.0 66 4.0 3 6.7 5 1.3 1 0.0 0
Colorectal 41 70.7 29 2.4 1 17.1 7 2.4 1 7.3 3
Pulmonary 26 38.5 10 11.5 3 38.5 10 3.8 1 7.7 2
Other gynecological
disorders 45 91.1 41 2.2 1 0.0 0 2.2 1 4.4 2
Others 56 51.8 29 7.1 4 23.2 13 8.9 5 8.9 5
TOTAL 520 52.1% 271 4.6% 24 17.1% 89 7.3% 38 18.8% 98
Non malignant
pathologies
Gastric 26 100.0 26 0.0 0 0.0 0 0.0 0 0.0 0
Pulmonary 16 93.8 15 6.3 1 0.0 0 0.0 0 0.0 0
Dialysis 30 86.7 26 13.3 4 0.0 0 0.0 0 0.0 0
Diabetes 34 85.3 29 0.0 0 14.7 5 0.0 0 0.0 0
Asthma 23 100.0 23 0.0 0 0.0 0 0.0 0 0.0 0
Cirrhosis 10 10.0 1 10.0 1 10.0 1 30.0 3 40.0 4
Hepatitis 41 78.0 32 0.0 0 17.1 7 2.4 1 2.4 1
Gynecological 31 93.5 29 3.2 1 0.0 0 0.0 0 3.2 1
TOTAL 211 85.8% 181 3.3% 7 6.2% 13 1.9% 4 2.8% 6

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PERFORMANCE Precision
Studies performed using VIDAS CA 125 II gave the
following results: Within-run reproducibility
Four samples and the control were tested n times in the
Measurement range
same run.
The measurement range of the VIDAS CA 125 II kit is: 4-
600 U/ml. Sample 1 2 3 4 5

Analytical detection limit n 30 30 29 29 28

Defined as the smallest concentration of OC 125 reactive Mean 33.5 29.4 94.3 231.7 566.1
antigenic determinants which is significantly different from (U/ml)
the zero concentration with a probability of 95%: < 4 U/ml.
CV % 3.4 3.6 3.1 3.7 3.9
Hook effect
No hook effect was found up to OC 125 reactive antigenic Between-run reproducibility
determinant concentrations of 200,000 U/ml. Four samples and the control were tested singly in n
different runs on the same VIDAS instrument.

Sample 1 2 3 4 5
n 30 30 29 30 29
Mean 33.8 28.6 93.1 225 561.3
(U/ml)
CV % 4.7 4.6 3.8 4.7 4.1

Comparison with other test methods

The concentration of OC 125 reactive antigenic determinants in a sample, determined using the kits from different
manufacturers, may vary depending on the test methods used. If the test method is changed and in the case of patient
monitoring, laboratories should confirm the concentrations previously found.
Correlation was established between VIDAS CA 125 II and a radioimmunoassay (X).
VIDAS CA 125 II = 1.13 X – 19.30 r = 0.97
(n = 137)

WASTE DISPOSAL 5. DAVIS H.M., ZURAWSKI V.R., BAST R.C. Jr et al.

Dispose of used or unused reagents and any other Characterization of the CA 125 antigen associated with
contaminated disposable materials following humanepithelial ovarian carcinomas. Cancer Research
1986 ; 46: 6143-6148.
procedures for infectious or potentially infectious
products. 6. HAGA Y., SAKAMOTO K., EGAMI H. et al. Evaluation of
serum CA 125 Values in healthy individuals and pregnant
It is the responsibility of each laboratory to handle
women. Am J Med Sci 1986 ; 292(1): 25-29.
waste and effluents produced according to their nature
and degree of hazardousness and to treat and dispose 7. KABAWAT S.E., BAST R.C. BHAN A.K., WELCH W.R.,
of them (or have them treated and disposed of) in KNAPP R.C., COLVIN R.B. Tissue distribution of a
accordance with any applicable regulations. coelomic-epithelium-related antigen recognized by the
monoclonal antibody OC 125. Int. J Gyn Path 1983 ; 2(3):
275-285.
LITERATURE REFERENCES
8. KABAWAT S.E., BAST R.C. Jr, KNAPP R.C. et al.
1. BARBIERI R.L., NILOFF J.M., BAST R.C. et al. Elevated Immunopathologic characterization of a monoclonal
serum concentrations of CA 125 in patients with advanced antibody that recognizes common surface antigens of
endometriosis. Fertil Steril 1986 ; 45(5): 630-634. human ovarian tumors of serous, endometrioid, and clear
cell types. Am J Clin Pathol 1983 ; 79(1): 98-104.
2. BAST R.C., FEENEY M., LAZARUS H. et al. Reactivity of
a monoclonal antibody with human ovarian carcinoma. J 9. NILOFF J.M., BAST R.C., SCHAETZL E.M. et al.
Clin Invest 1981 ; 68: 1331-1337. Predictive value of CA 125 antigen levels in second-look
procedures for ovarian cancer. Am J Obstet Gynecol
3. BAST R.C., KLUG T.L., ST JOHN E. et al. A radio
1985 ; 151: 981-986.
immunoassay using a monoclonal antibody to monitor the
course of epthelial ovarian Cancer. N Engl J Med 1983 ; 10. PITTAWAY D.E. CA 125 in women with Endometriosis.
309(15): 883-887. Obstet Gynecol Clin North Am 1989 ; 16(1): 237-252.
4. BERGMANN J.F., BEAUGRAND M., LABADIE H. et al. 11. SCHILTUIS M.S., AALDERS J.G., BOUMA J. et al. Serum
CA 125 (Ovarian tumor associated antigen) in ascitic liver CA 125 levels in epithelial ovarian cancer: Relation with
diseases. Clin Chim Acta 1986 ; 155: 163-166. findings at second-look operations and their role in the
detection of tumour recurrence. Br J Obstet Gynecol
1987 ; 94: 202-207.
12. WATANABE N. and al. In vitro effect of contrast agents
during immunoradiometric assay for tumor-associated
antigens. Nuclear Medicine Communications, 1998, 19,
63-70.

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INDEX OF SYMBOLS WARRANTY

Symbol Meaning bioMérieux disclaims all warranties, express or implied,
including any implied warranties of MERCHANTABILITY
AND FITNESS FOR A PARTICULAR USE. bioMérieux
Catalog number
shall not be liable for any incidental or consequential
damages. IN NO EVENT SHALL BIOMERIEUX’S
In Vitro Diagnostic Medical Device LIABLITY TO CUSTOMER UNDER ANY CLAIM
EXCEED A REFUND OF THE AMOUNT PAID TO
BIOMERIEUX FOR THE PRODUCT OR SERVICE
Manufacturer WHICH IS THE SUBJECT OF THE CLAIM.

Temperature limit

Use by date

Batch code

Consult Instructions for Use

Contains sufficient for <n> tests

Date of manufacture

REVISION HISTORY
Change type categories :
N/A Not applicable (First publication)
Correction Correction of documentation anomalies
Technical change Addition, revision and/or removal of information related to the product
Administrative Implementation of non-technical changes noticeable to the user
Note: Minor typographical, grammar, and formatting changes are not included in the
revision history.

Release
Part Number Change Type Change Summary
date
INDEX OF SYMBOLS
Administrative REVISION HISTORY
2015/01 08587H CONTENT OF THE KIT (30 TESTS) – RECONSTITUTION
Technical OF REAGENTS
WARNINGS AND PRECAUTIONS
INSTRUCTIONS FOR USE

BIOMERIEUX, the blue logo, SPR and VIDAS are used, pending and/or registered trademarks belonging to bioMérieux, or one of its
subsidiaries, or one of its companies.
CA 125 II is a trademark belonging to Fujirebio Diagnostics Inc.
Any other name or trademark is the property of its respective owner.

673 620 399 RCS LYON

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