BIOMÉRIEUX

30 429 08814 J - en - 2017/10

VIDAS® CA 15-3®(153)
VIDAS CA 15-3 is an automated quantitative test for use on the VIDAS family instruments for the quantitative
measurement of CA 15-3 levels in human serum or plasma (lithium heparin or EDTA) using the ELFA technique
(Enzyme Linked Fluorescent Assay).

SUMMARY AND EXPLANATION PRINCIPLE

The VIDAS CA 15-3 test uses two monoclonal antibodies The assay principle combines a 2-step enzyme
(115D8 and DF3) which react with a circulating CA 15-3, immunoassay sandwich method with a final fluorescent
antigen expressed in human breast cancer cell. The detection (ELFA).
®
monoclonal 115D8 antibody directed against human milk The Solid Phase Receptacle (SPR ) serves as the solid
fat globule membranes, and the monoclonal DF3 antibody phase as well as the pipetting device for the assay.
directed against an enriched fraction of human metastatic Reagents for the assay are ready-to-use and pre-
breast cancer membrane, react with the epitopes dispensed in the sealed reagent strips.
expressed by a family of high molecular weight All of the assay steps are performed automatically by the
glycoproteins known as polymorphic epithelial mucins (10, instrument. The reaction medium is cycled in and out of
11, 13, 18). the SPR several times.
An increase in the concentration of CA 15-3 is frequently
found in breast cancer, as in certain other cancers, but The sample is cycled in and out of the SPR several times.
This operation enables the 115D8 fixed onto the interior
also in certain non-cancerous pathologies. The CA 15-3
concentration decreases after therapy and increases in wall of the SPR to capture the reactive antigenic
determinants present in the sample. Unbound
cases of relapse, residual disease and metastasis.
The VIDAS CA 15-3 test is used for the monitoring of components are eliminated during the washing steps.
Alkaline phosphatase-labeled DF3 antibody is then
therapy for patients with diagnosed malignant tumors and
to evaluate the evolution of the pathology. A decrease in incubated in the SPR where it binds with the DF3 reactive
antigenic determinants. Unbound conjugate is then
the CA 15-3 concentration can indicate a positive
response to therapy and therefore good prognosis. A eliminated during the washing steps.
During the final detection step, the substrate (4-Methyl-
constant increase in CA 15-3 concentration often reflects
evolution of the tumor and a poor response to therapy (1- umbelliferyl phosphate) is cycled in and out of the SPR.
The conjugate enzyme catalyzes the hydrolysis of this
9, 12, 14-17, 19).
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 CA 15-3 present in the sample.
At the end of the assay, the results are automatically
calculated by the instrument in relation to the calibration
curve stored in memory, and then printed out.
CONTENT OF THE KIT (30 TESTS):
30 CA 15-3 strips STR Ready-to-use.
30 CA 15-3 SPRs SPR Ready-to-use.
1 x 30 Interior of SPR coated with monoclonal 115D8 antibodies (mouse).
CA 15-3 Control C1 Ready-to-use.
1 x 1 ml (liquid) Bovine albumin + DF3 reactive antigenic determinants (human origin) + 0.9 g/l
sodium azide.
MLE data indicate the confidence interval in U/mL ("Control C1 Dose Value
Range").
CA 15-3 Calibrator S1 Ready-to-use.
1 x 1.5 ml (liquid) Bovine albumin + DF3 reactive antigenic determinants (human origin) + 0.9 g/l
sodium azide.
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 15-3 Diluent R1 Ready-to-use.
1 x 5 ml (liquid) Calf serum + 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 codes printed on the box label.
1 Clip seal
1 Package Insert provided in the kit or downloadable from www.biomerieux.com/techlib.

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The SPR The Reagent Strip

The SPR is coated during production with monoclonal The strip consists of 10 wells covered with a labeled, foil
115D8 antibodies (mouse). Each SPR is identified by the seal. The label comprises a bar code which mainly
"153" code. Only remove the required number of SPRs indicates the assay code, kit lot number and expiration
from the pouch. Carefully reseal the pouch after date. The foil of the first well is perforated to facilitate the
opening using the clip seal provided with the kit. 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 15-3 Reagent Strip
Wells Reagents
1 Sample well.
2-3-4 Empty wells.
5 Conjugate: Alkaline phosphatase-labeled DF3 antibody + 0.9 g/l sodium azide
(400 µl).
6-7 Wash buffer: Tris (0.1 mol/l, pH 7.4) + NaCl (0.1 mol/l) + Tween (0.05%) + 0.9 g/l
sodium azide (600 µl).
8 Diluent: Tris (0.1 mol/l) + NaCl (0.1 mol/l) + calf serum (5%) + 0.9 g/l sodium azide
(400 µl).
9 Wash buffer: Tris (0.1 mol/l, pH 7.4) + NaCl (0.1 mol/l) + Tween (0.05%) +
0.9 g/l sodium azide (600 µl).
10 Cuvette with substrate: 4-Methyl-umbelliferyl phosphate (0.6 mmol/l) +
diethanolamine (DEA*) (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.
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 • This kit contains products of animal origin. Certified
NOT PROVIDED knowledge of the origin and/or sanitary state of the
- Pipette with disposable tip to dispense 100 µl. animals does not totally guarantee the absence of
- Powderless, disposable gloves. transmissible pathogenic agents. It is therefore
- For other specific materials and disposables, please recommended that these products be treated as
refer to the Instrument User’s Manual. potentially infectious and handled observing the usual
- Instrument of the VIDAS family. safety precautions (do not ingest or inhale).
• Do not use the SPRs if the pouch is pierced.
WARNINGS AND PRECAUTIONS
• Do not use visibly deteriorated SPRs (damaged foil or
• For in vitro diagnostic use only. plastic).
• For professional use only. • Do not use reagents after the expiration date indicated
• This kit contains products of human origin. No on the label.
known analysis method can totally guarantee the • Do not mix reagents (or disposables) from different lots.
absence of transmissible pathogenic agents. It is • Use powderless gloves, as powder has been reported
therefore recommended that these products be to cause false results for certain enzyme immunoassay
treated as potentially infectious and handled tests.
observing the usual safety precautions (see • Kit reagents contain sodium azide which can react with
Laboratory Biosafety Manual - WHO - Geneva - lead or copper plumbing to form explosive metal azides.
latest edition). If any liquid containing sodium azide is disposed of in
the plumbing system, drains should be flushed with
water to avoid build-up.
• The optical cuvette with substrate (well 10) contains an
irritant agent (6.6% diethanolamine). Refer to the hazard
statements "H" and the precautionary statements "P"
above.

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• Spills should be wiped up thoroughly after treatment Calibration

with liquid detergent or a solution of household bleach
Calibration, using the calibrator provided in the kit, must
containing at least 0.5% sodium hypochlorite. See the
be performed upon receipt of a new lot of reagents after
User’s Manual for cleaning spills on or in the instrument.
the master lot data have been entered. Calibration should
Do not autoclave solutions containing bleach.
then be performed every 14 days. This operation provides
• The instrument should be regularly cleaned and
instrument-specific calibration curves and compensates
decontaminated (see the User’s Manual).
for possible minor variations in assay signal throughout
the shelf-life of the kit.
STORAGE CONDITIONS The calibrator, identified by S1, must be tested in
• Store the VIDAS CA 15-3 kit at 2-8°C. duplicate (see User’s Manual).
• Do not freeze reagents. The calibrator value must be within the set RFV "Relative
• Store all unused reagents at 2-8°C. Fluorescence Value" range. If this is not the case,
• After opening the kit, check that the SPR pouch is recalibrate.
correctly sealed and undamaged. If not, do not use the
SPRs. Procedure
• To maintain the stability of the SPRs, carefully 1. Only remove the required reagents from the
reseal the pouch after use with the desiccant inside, refrigerator and allow them to come to room
using the clip seal provided, and return the temperature for at least 30 minutes.
complete kit to 2-8°C. 2. Use one "153" strip and one "153" SPR from the kit for
• If stored according to the recommended conditions, all each sample, control or calibrator to be tested. Make
components are stable until the expiration date indicated sure the storage pouch has been carefully
on the label. resealed with the clip seal after the required SPRs
have been removed.
SPECIMENS 3. The test is identified by the "153" code on the
Specimen type and collection: instrument. The calibrator must be identified by "S1",
and tested in duplicate. If the control is to be tested, it
Human serum or plasma (lithium heparin or EDTA).
should be identified by "C1".
It is recommended that each laboratory checks the
compatibility of collection tubes used. 4. Mix the calibrator, control and samples using a vortex-
The use of heat inactivated sera has not been validated. type mixer (for serum or plasma separated from the
None of the following factors have been found to pellet).
significantly influence this assay. 5. For this test, the calibrator, control, and sample
- hemolysis (after spiking samples with hemoglobin: 0 to test portion is 100 µl.
300 µmol/l (monomer)), 6. Insert the "153" SPRs and "153" strips into the
- lipemia (after spiking samples with lipids: 0 to 10 mg/ml instrument. Check to make sure the color labels with
equivalent in triglycerides), the assay code on the SPRs and the Reagent Strips
- bilirubinemia (after spiking samples with bilirubin: 0 to match.
470 µmol/l). 7. Initiate the assay as directed in the User’s Manual. All
However, it is recommended not to use samples that are the assay steps are performed automatically by the
clearly hemolyzed, lipemic or icteric and, if possible, to instrument.
collect a new sample. 8. Reclose the vials and return them to 2–8°C after
Specimen stability: pipetting.
Samples can be stored at 2-8°C in stoppered tubes for up 9. The assay will be completed within approximately
to 48 hours; if longer storage is required, freeze the sera 60 minutes. After the assay is completed, remove the
or plasma at -25 ± 6°C. SPRs and strips from the instrument.
Avoid successive freezing and thawing. 10. Dispose of the used SPRs and strips into an
A study performed on frozen samples over a period of appropriate recipient.
2 months, showed that the quality of results is not
affected. RESULTS AND INTERPRETATION
Once the assay is completed, results are analyzed
INSTRUCTIONS FOR USE automatically by the computer. Fluorescence is measured
For complete instructions, see the User’s Manual. twice in the Reagent Strip’s reading cuvette for each
sample tested. The first reading is a background reading
Reading Master lot data of the substrate cuvette before the SPR is introduced into
Before each new lot of reagents is used, enter the the substrate. The second reading is taken after
specifications (or factory master data) into the instrument incubating the substrate with the enzyme remaining on
using the master lot entry (MLE) data. the interior of the SPR. The RFV (Relative Fluorescence
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.
Note: the master lot data need only be entered once The results are automatically calculated by the instrument
for each lot. using calibration curves which are stored by the
It is possible to enter MLE data manually or instrument (4-parameter logistic model). The
automatically depending on the instrument (refer to the concentrations are expressed in U/ml.
User’s Manual).

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Samples with CA 15-3 titers > 400 U/ml should be RANGE OF EXPECTED VALUES
reassayed after maximum dilution by 1/10 in CA 15-3 These figures are given as a guide; it is recommended
diluent. that each laboratory establish its own reference values
If the dilution factor has not been entered when the Work from a rigorously selected population.
List was created (see User’s Manual), multiply the result The following results were obtained using 543 samples
by the dilution factor to obtain the sample concentration.
collected from 134 healthy subjects, 347 subjects with a
Interpretation of test results should be made taking into
consideration the patient’s history, and the results of any diagnosed malignant pathology, and 62 subjects with a
other tests performed. non-malignant affection:

No. Percentage (%) in relation to

QUALITY CONTROL
of the zone of values in U/ml
A control is included in each VIDAS CA 15-3 kit.
This control must be performed immediately after opening subjects < 30 30-60 60- > 120
a new kit to ensure that reagent performance has not 120 U/ml
U/ml U/ml
been altered. Each calibration must also be checked U/ml
using this control. The instrument will only be able to
check the control value if it is identified by C1.
Results cannot be validated if the control value deviates Healthy female
from the expected values. subjects
Note < 50 years 69 99 1 0 0
It is the responsibility of the user to perform Quality >= 50 years 65 97 3 0 0
Control in accordance with any local applicable
Malignant
regulations.
pathologies
LIMITATIONS OF THE METHOD
Breast
Samples collected from patients receiving mouse
monoclonal antibody preparations for diagnostic or Stage I 25 84 16 0 0
therapeutic purposes may contain human anti-mouse Stage II 25 68 24 8 0
antibodies (HAMA). These samples may give falsely high
Stage III 25 56 24 12 8
or low concentrations when tested with kits containing
mouse antibodies. Stage IV 25 20 12 28 40
The VIDAS CA 15-3 assay is based on the use of Colorectal 49 86 10 4 0
monoclonal DF3 and 115D8 antibodies which are
supplied exclusively by Fujirebio Diagnostics Inc., their Pulmonary 46 65 26 7 2
distributors and licensed organizations. Methods using Ovarian 56 66 32 0 2
antibodies other than DF3 and 115D8 can give different 0
Pancreatic 19 69 26 5
clinical results.
Results from this test should be interpreted as part of a Thyroid 27 93 7 0 0
complete clinical profile and in conjunction with other Other pathologies 50 80 20 0 0
diagnostic techniques. The VIDAS CA 15-3 assay cannot
be used as a screening test for cancer. Gynecological
Interference may be encountered with certain sera Non malignant
containing antibodies directed against reagent pathologies
components. For this reason, assay results should be
interpreted taking into consideration the patient history, Renal 26 77 19 4 0
and the results of any other tests performed. Hepatic 29 97 3 0 0
It is not advisable to perform CA 15-3 assays on patients
Others 7 100 0 0 0
who have received a contrast agent in the previous 24
hours [20].
PERFORMANCE
Studies performed using VIDAS CA 15-3 gave the
following results:
Measurement range
The measurement range of the VIDAS CA 15-3 kit
extends from 2 to 400 U/ml.
Analytical detection limit
Defined as the smallest concentration of DF3 reactive
antigenic determinants which is significantly different from
the zero concentration with a probability of 95%: 2 U/ml.
Hook effect
No hook effect was found up to DF3 reactive antigenic
determinant concentrations of 38,000 U/ml.

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Precision Between-run reproducibility:

Within-run reproducibility:
5 samples were tested in 29 different runs on the same
5 samples were tested 30 times in a same run. VIDAS instrument.

Sample 1 2 3 4 5 Sample 1 2 3 4 5
Mean (U/ml) 5.3 10.9 22.0 105.1 374.9 Mean (U/ml) 6.1 11.4 22.5 100.4 354.8

CV % 3.6 3.2 3.5 2.7 3.4 CV % 5.4 4.8 4.8 4.0 4.9

Accuracy
Dilution test
The serum matrix of the sample can influence the results of the dilution test. When printing out results, it is
recommended to indicate the level of dilution used.
2 samples were diluted in the CA 15-3 diluent and tested singly in 3 runs. The ratio of the mean concentration measured
over the expected concentration is expressed as a mean recovery percentage.
Sample Dilution factor Expected concentration Mean measured Mean recovery
(U/ml) concentration percentage
(U/ml) (%)
1 1/1 186.2 186.2 100
1/2 93.1 96.8 104
1/4 46.5 49.6 107
1/8 23.3 25.2 108
1/16 11.6 13.5 116
2 1/1 266.5 266.5 100
1/2 133.2 152.4 114
1/4 66.6 85.0 128
1/8 33.3 43.3 130
1/16 16.7 22.9 137

Comparison with other test methods WASTE DISPOSAL

The concentration of DF3 reactive antigenic determinants Dispose of used or unused reagents as well as any other
may vary in a sample determined using the kits from contaminated disposable materials following procedures
different manufacturers, depending on the test method for infectious or potentially infectious products.
used. It is the responsibility of each laboratory to handle waste
If the test method is changed and in the case of patient and effluents produced according to their type and degree
monitoring, laboratories should confirm the concentrations of hazardousness and to treat and dispose of them (or
previously found. have them treated and disposed of) in accordance with
Correlation was established between the VIDAS CA 15-3 any applicable regulations.
kit and a radioimmunoassay method.
VIDAS = 0.89 IRMA technique + 1.1
n = 171 r = 0.997.

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LITERATURE REFERENCES INDEX OF SYMBOLS

1. BON GG., KENEMANS P., YEDEMAC A., et al. Clinical
Relevance of the Tumor Marker CA 15-3 in the Management Symbol Meaning
of Cancer Patients. In: From Clone To Clinic. The
Netherlands ; Kluwer Academics Publishers, 1990: 111-22. Catalog number
2. COLOMER R., RUIBAL A., GENOLIA J. et al. Circulating CA
15-3 Levels in the Postsurgical Follow-up of Breast Cancer
Patients and in non-malignant Diseases. Breast Cancer Res. In Vitro Diagnostic Medical Device
Treat. 1989 ; 13: 123-33.
3. COLOMER R., RUIBAL A., GENOLLA J., SALVADOR L.
Circulating CA 15-3 antigen levels in non-mammary Manufacturer
malignancies. Br. J. Cancer 1989 ; 59: 283-6.
4. COLOMER R., RUIVAL A., NAVARRO M., ENCABO G.,
SOLE L.A. Circulating CA 15-3 levels in breast cancer. Our Temperature limit
present experience. In. J. Biol. Markers. 1987 ; 1: 89-92.
5. COLOMER R., RUIBAL A., SALVADOR L. Circulating Tumor
Marker Levels in Advanced Breast Carcinoma Correlate with Use by date
the Extent of Metastatic Disease. Cancer 1989 ; 64: 1674-
1681.
6. GERAGHTY JG., COVENEY MB., SHERRY F., et al. CA 15- Batch code
3 in patients with locoregional and metastatic breast
carcinoma. Cancer 1992 ; 70: 2831-2834. Consult Instructions for Use
7. GION M., MIONE R., DITTADI S., FASAN S., PALLINI A.,
BRUSCAGNIN G. Evaluation of CA 15-3 serum levels in Contains sufficient for <n> tests
breast cancer patients. J. Nucl. Med. Al. Sciences 1986 ;
30: 29-35.
8. GION M., MIONE R., NASCIMBE O., et I. The tumor
associated antigen CA 15-3 in primary breast cancer.
Caution, consult accompanying
Evaluation of 667 cases. Br. J. Cancer 1191 ; 63: 809-813. documents
9. HAYES DF., SEKINE H., OHNO T., et al. Use of Murine
Monoclonal antibody for Detection of Circulating Plasma DF3
Antigen Levels in Breast Cancer Patients. J.C Invest. 1985 ; Date of manufacture
75: 1671-1678.
10. HILKENS J., BUIJS F., HILGERS J. et al. Monoclonal
Antibodies Against Human Milk-Fat Globule Membranes LIMITED WARRANTY
Detecting Differentiation Antigens of the Mammary Gland and bioMérieux warrants the performance of the product for its
its Tumors. Int. J. Cancer. 1984 ; 34: 197-206. stated intended use provided that all procedures for
11. HILKENS J., HILGERS J., BUIJS F. et al. Monoclonal usage, storage and handling, shelf life (when applicable),
Antibodies Against Human Milk-Fat Globule Membranes and precautions are strictly followed as detailed in the
Useful in Carcinoma Research. Prot. Biol. Fluids. 1984 ; 31:
instructions for use (IFU).
1013-6.
Except as expressly set forth above, bioMerieux hereby
12. HILKENS J., KROEZEN V., BONFRER JM., et al. MAM-6
Antigen. A New marker for Breast Cancer Monitoring. Cancer
disclaims all warranties, including any implied warranties
Res. 1986 ; 46: 2582-7. of merchantability and fitness for a particular purpose or
13. KUFE D., INGHIRAMI G., ABE M., et al. Differential use, and disclaims all liability, whether direct, indirect or
Reactivity of a Novel Monoclonal Antibody (DF3) with Human consequential, for any use of the reagent, software,
Malignant vs Benign Breast Tumors. Hybridoma 1984 ; 3: instrument and disposables (the “System”) other than as
223-32. set forth in the IFU.
14. PONS-ANICET DM., KREBS BP., MIRA R., et al. Value of
CA 15-3 in the follow-up of breast cancer patients. Br.J.
Cancer 1987 ; 55: 567-9.
15. ROBERTSON JFR., PEARSON D., PRICE MR., et al.
Assessment of four monoclonal antibodies as serum markers
in breast cancer. Eur. J. Cancer. 1990 ; 26: 1127-1132.
16. SAFI F., KOHLER I., RÖTTINGER E., BEGER H-G. The
value of the Tumor Marker CA 15-3 in Diagnosing and
Monitoring Breast Cancer. 1991 ; 68: 574-582.
17. SILVER H., ARCHIBALD B-L., RAGAZ J., COLDMAN A. A
relative operating characteristic analysis and group modeling
for Tumor Markers: comparison of CA 15-3,
carcinoembryogenic Antigen, and Mucin-like carcinoma
associated antigen in breast carcinoma. Cancer Research
1991 ; 51: 1904-1909.
18. TAYLOR-PAPAMIDITRIOU J., GENDLER S. Molecular
Aspects of Mucins. Cancer Rev. 1988 ; 11-12: 11-24.
19. TONDINI C., HAYES DF., GELMAN R., et al. Comparison of
CA 15-3 and carcinoembryonic antigen in monitoring the
clinical course of patients with metastatic breast cancer.
Cancer Research 1988 ; 48: 4107-4112.
20. 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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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 08814 I
CONTENT OF THE KIT (30 TESTS)
Technical WARNINGS AND PRECAUTIONS
INSTRUCTIONS FOR USE

Administrative LIMITED WARRANTY

2017/10 08814 J
STORAGE CONDITIONS
Technical
RANGE OF EXPECTED VALUES

BIOMERIEUX, the BIOMERIEUX 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 15-3 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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