30 421 07709 J - en - 2015/01

VIDAS® CK-MB (CKMB)

VIDAS CK-MB is an automated quantitative test for use on the VIDAS family instruments for the quantitative
measurement of the MB isoenzyme of human creatine kinase in human serum or plasma (lithium heparin or EDTA),
using the ELFA technique (Enzyme Linked Fluorescent Assay).

SUMMARY AND EXPLANATION PRINCIPLE

Creatine kinase (CK) is a key enzyme of muscle The assay principle combines the enzyme immunoassay
metabolism. It is present in all tissues, but in variable sandwich method with a final fluorescent detection (ELFA)
concentrations and in different isoforms (CK-MM, CK-MB (5, 6, 7).
®
and CK-BB) (1). In skeletal muscle, creatine kinase is The Solid Phase Receptacle (SPR ) serves as the solid
essentially found in the MM isoform (1). The BB isoform is phase as well as the pipetting device for the assay.
found predominantly in the brain and the MB fraction is Reagents for the assay are ready-to-use and pre-
found principally in the heart tissue (1, 2). dispensed in the sealed reagent strips.
In the absence of any major muscular trauma, an All of the assay steps are performed automatically by the
elevation of serum CK-MB concentration indicates instrument. The reaction medium is cycled in and out of
necrotic injury to the heart occurring as a consequence of the SPR several times.
myocardial infarction. Generally, CK-MB is detectable five A pre-wash step prepares the SPR for the reaction. The
hours after the onset of chest pains and peak sample is then transferred into the well containing the
concentrations are often reached eleven to eighteen serum diluent. The sample/serum diluent mixture is cycled
hours after the infarction (3, 4). in and out of the SPR. Unbound components are
There exists, however, a certain number of other eliminated during the washing steps. The conjugate
situations, pathological or otherwise, in which an elevation (alkaline phosphatase-labeled anti-CK-MM antibody
of CK-MB can be observed, such as Duchenne’s disease, fragment) is cycled in and out of the SPR. Unbound
muscular hyperactivity, hypo- and hyperthermia, and components are eliminated during the washing steps.
myocarditis (2). During the final detection step, the substrate (4-Methyl-
The VIDAS CK-MB assay aids in diagnosing acute umbelliferyl phosphate) is cycled in and out of the SPR.
coronary syndromes. 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 CK-MB 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.

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CONTENT OF THE KIT (30 TESTS) - RECONSTITUTION OF REAGENTS

30 CKMB strips STR Ready-to-use.
®
30 CKMB SPRs SPR Ready-to-use.
1 x 30 Interior of SPRs coated with monoclonal anti-CK-MB immunoglobulins (mouse).
CKMB control C1 Reconstitute with 3 ml of reconstitution solvent. Wait for 10 to 15 minutes, then
1 x 3 ml (lyophilized) mix. After reconstitution, stable for 24 hours at 2-8°C: In aliquots and frozen at -
25  6°C, stable until the expiration date on the kit.
Avoid successive freeze/thaw cycles.
Thaw at room temperature (18-25°C).
Protein base (BSA) + chemical stabilizers + human CK-MB.
MLE data indicate the confidence interval in ng/mL ("Control C1 Dose Value
Range").
CKMB calibrator S1 Reconstitute with 3 ml of reconstitution solvent. Wait for 10 to 15 minutes, then
1 x 3 ml (lyophilized) mix. After reconstitution, stable for 24 hours at 2-8°C. In aliquots and frozen at -
25  6°C, stable until the expiration date on the kit.
Avoid successive freeze/thaw cycles. Thaw at room temperature (18-25°C).
Protein base (BSA) + chemical stabilizers + human CK-MB.
MLE data indicate the concentration in ng/mL (“Calibrator (S1) Dose Value") and
the confidence interval in "Relative Fluorescence Value ("Calibrator (S1) RFV
Range”).
Reconstitution solvent R1 Ready to use.
1 x 20 ml (liquid) Demineralized water + 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 Package Insert provided in the kit or downloadable from www.biomerieux.com/techlib.

The SPR The strip

The SPR is coated during production with monoclonal The strip consists of 10 wells covered with a labeled foil
anti-CK-MB immunoglobulins (mouse). Each SPR is seal. The label comprises a bar code which mainly
identified by the CKMB code. Only remove the required indicates the assay code, kit lot number and expiration
number of SPRs from the pouch and carefully reseal the date. The foil of the first well is perforated to facilitate the
pouch after opening. introduction of the sample. The last well of each strip is a
cuvette in which the fluorimetric reading is performed. The
wells in the center section contain the various reagents
required for the assay.

Description of the CKMB strip

Wells Reagents
1 Sample.
2 Serum diluent: Tris-NaCl (0.05 mol/l, pH 7.4), protein and chemical stabilizers,
0.9 g/l sodium azide (300 µl).
3 Wash buffer: Tris-NaCl (0.05 mol/l, pH 7.4), protein stabilizers,
0.9 g/l sodium azide (600 µl).
4-5-7-8 Wash buffer: Tris-NaCl (0.05 mol/l, pH 7.4), surfactant,
0.9 g/l sodium azide (600 µl).
6 Conjugate: alkaline phosphatase-labeled anti-CK-MM polyclonal antibody Fab’
fragment (goat), Tris-NaCl buffer (0.05 mol/l, pH 6), protein stabilizers,
0.9 g/l sodium azide (400 µl).
9 Empty well.
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

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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 STORAGE CONDITIONS
NOT PROVIDED  Store the VIDAS CK-MB kit at 2-8°C.
- Pipette with disposable tip to dispense 3 ml and 250 µl.  Do not freeze reagents, with the exception of
- Powderless, disposable gloves. calibrators and controls after reconstitution.
- For other specific materials and disposables, please  Store all unused reagents at 2-8°C.
refer to the Instrument User’s Manual.  After opening the kit, check that the SPR pouch is
- Instrument of the VIDAS family. correctly sealed and undamaged. If not, do not use the
SPRs.
WARNINGS AND PRECAUTIONS  Carefully reseal the pouch with the desiccant inside
 For in vitro diagnostic use only. after use to maintain the stability of the SPRs and
 For professional use only. return the complete kit to 2-8°C.
 This kit contains products of human origin. No  If stored according to the recommended conditions, all
known analysis method can totally guarantee the components are stable until the expiration date indicated
absence of transmissible pathogenic agents. It is on the label. Refer to the kit composition table for
therefore recommended that these products be special storage conditions.
treated as potentially infectious, and handled
observing the usual safety precautions (see SPECIMENS
Laboratory Biosafety Manual - WHO - Geneva - Specimen type and collection
latest edition).
Serum or plasma (lithium heparin or EDTA).
 This kit contains products of animal origin. Certified
It is recommended that each laboratory checks the
knowledge of the origin and/or sanitary state of the
compatibility of collection tubes used.
animals does not totally guarantee the absence of
None of the following factors have been found to
transmissible pathogenic agents. It is therefore
recommended that these products be treated as significantly influence this assay:
potentially infectious and handled observing the usual - hemolysis (after spiking samples with hemoglobin: 0 to
safety precautions (do not ingest or inhale). 5 g/l (monomer)),
 Do not use SPRs if the pouch is pierced. - lipemia (after spiking samples with lipids: 0 to
34.2 mmol/l equivalent in trigycerides),
 Do not use visibly deteriorated STRs (damaged foil or
- bilirubinemia (after spiking samples (naturally icteric
plastic).
plasma) with bilirubin: 0 to 850 µmol/l).
 Do not use reagents after the expiration date indicated
on the label. However, it is recommended not to use samples that are
clearly hemolyzed, lipemic or icteric and, if possible, to
 Do not mix reagents (or disposables) from different lots.
collect a new sample.
 Use powderless gloves as powder has been reported
to cause false results for certain enzyme immunoassay Specimen stability
tests (8). Samples can be stored at 2-8°C in stoppered tubes for up
 Kit reagents contain sodium azide which can react with to 48 hours.
lead or copper plumbing to form explosive metal azides. If longer storage is required, freeze the sera or plasma at
If any liquid containing sodium azide is disposed of in at -25  6 C.
the plumbing system, drains should be flushed with Avoid successive freezing and thawing.
water to avoid build-up. Thaw at room temperature (18-25°C).
 The optical cuvette with substrate (well 10) contains an A study performed on frozen samples over a period of
irritant agent (6.6% diethanolamine). Refer to the hazard 5 months showed that the quality of results is not affected.
statements "H" and the precautionary statements "P"
above. INSTRUCTIONS FOR USE
 Spills should be wiped up thoroughly after treatment For complete instructions, see the User’s Manual.
with liquid detergent or a solution of household bleach
containing at least 0.5% sodium hypochlorite. See the Reading Master lot data
User's Manual for cleaning spills on or in the instrument. Before each new lot of reagents is used, enter the
Do not autoclave solutions containing bleach. specifications (or factory master data) into the instrument
 The instrument should be regularly cleaned and using the master lot entry (MLE) data.
decontaminated (see the User's Manual). If this operation is not performed before initiating the
tests, the instrument will not be able to print results.
Note: the master lot data need only be entered once
for each lot.
It is possible to enter MLE data manually or
automatically depending on the instrument (refer to the
User’s Manual).

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Calibration The results are automatically calculated by the instrument

Calibration, using the calibrator provided in the kit, must using calibration curves which are stored in memory
be performed each time a new lot of reagents is opened, (4-parameter logistic model) and are expressed in ng/ml.
after the master lot data have been entered. Calibration The measurement range for VIDAS CK-MB is
should then be performed every 14 days. This operation 0.8-300 ng/ml. Samples with CK-MB concentrations
provides instrument-specific calibration curves and greater than 300 ng/ml must be retested after dilution 1/4
compensates for possible minor variations in assay signal in human serum with a CK-MB concentration < 0.8 ng/ml.
throughout the shelf-life of the kit. If the dilution factor has not been entered when the Work
The calibrator, identified by S1, must be tested in List was created (see User's Manual), multiply the result
duplicate (see User's Manual). The calibrator value must by the dilution factor to obtain the sample concentration.
be within the set RFV (Relative Fluorescence Value). If Interpretation of test results should be made taking into
this is not the case: recalibrate. consideration the patient's history, and the results of any
other tests performed.
Test procedure
QUALITY CONTROL
1. Only remove the reagents required from the
refrigerator. A positive control is included in each VIDAS CK-MB kit.
This control must be performed immediately after opening
2. Use one “CKMB” strip and one “CKMB” SPR for each a new kit to check that reagent performance has not been
sample, control or calibrator to be tested. Make sure altered. Each calibration must also be checked using this
the storage pouch has been carefully resealed control. The instrument will only be able to check the
after the required SPRs have been removed. control value if it is identified by C1.
3. The test is identified by the “CKMB” code on the Results cannot be validated if the control value deviates
instrument. The calibrator must be identified by "S1", from the expected values.
and tested in duplicate. If the control is to be tested, it
should be identified by "C1". Note
4. If necessary, clarify samples by centrifugation. It is the responsibility of the user to perform Quality
Control in accordance with any local applicable
5. Mix the calibrator, controls and samples using a regulations.
vortex-type mixer (for serum or plasma separated from
the pellet).
LIMITATIONS OF THE METHOD
6. For this test, the calibrator, control, and sample Interference may be encountered with certain sera
test portion is 250 µl. containing antibodies directed against reagent
7. Insert the “CKMB” SPRs and “CKMB” strips into the components. For this reason, assay results should be
instrument. Check to make sure the color labels with interpreted taking into consideration the patient history,
the assay code on the SPRs and the Reagent Strips and the results of any other tests performed.
match.
8. Initiate the assay as directed in the User's Manual. All RANGE OF EXPECTED VALUES
the assay steps are performed automatically by the These figures are given as a guide. It is recommended
instrument. that each laboratory establishes its own reference values
9. Reclose the vials and return them to the required from a rigorously selected population.
temperature after pipetting. Depending on the patients tested, differences in the
10. The assay will be completed within approximately expected values may be found
30 minutes. After the assay is completed, remove the The assay of ponderal CK-MB can be completed with the
SPRs and strips from the instrument. assay of total CK using a relative index (RI):
11. Dispose of the used SPRs and reagent strips into an
CK-MB (ng/ml)
appropriate recipient.
RI (%) = ______________________________ x 100
Total CK (U/l)
RESULTS AND INTERPRETATION
Once the assay is completed, results are analyzed It is recommended that each laboratory establishes its
automatically by the computer. Fluorescence is measured own reference values for the calculation of the relative
twice in the Reagent Strip’s reading cuvette for each index (RI) due to the differences between the total CK
sample tested. The first reading is a background reading methods and the levels of CK-MB obtained for different
of the substrate cuvette before the SPR is introduced into populations. As an indication, for a reference population of
the substrate. The second reading is taken after 119 blood donors, the upper limit of the 95% reference
incubating the substrate with the enzyme remaining on range of CK-MB was determined as 5.1 ng/ml.
the interior of the SPR. The RFV (Relative Fluorescence
Value) is calculated by subtracting the background
reading from the final result. This calculation appears on
the result sheet.

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PERFORMANCE Precision
Studies performed with the VIDAS CK-MB assay gave the Within-run reproducibility:
following results:
Three samples were tested 30 times in the same run.
Measurement range
The measurement range of the VIDAS CK-MB reagent is Samples E1 E2 E3
0.8-300 ng/ml. Number of measurements 30 30 30
Analytical detection limit Mean concentration (ng/ml) 3.73 11.94 94.79
Defined as the smallest concentration of CK-MB which is CV (%) 6.16 3.42 3.29
significantly different from the zero concentration with a
probability of 95%: 0.8 ng/ml. Between-run reproducibility:
Hook effect Three samples were tested singly in 23 different runs on
No hook effect was observed up to CK-MB concentrations the same VIDAS instrument.
of 10,000 ng/ml.
Samples E1 E2 E3
Number of measurements 23 23 23
Mean concentration (ng/ml) 2.89 12.49 114.34
CV (%) 5.13 4.95 3.13

Specificity
No cross-reactivitiy was observed with CK-MM and CK-
BB for tested concentrations of up to 10,000 ng/ml.
Accuracy

Dilution test

3 human serum samples were diluted in a weak serum. The ratio of the mean concentration measured over the expected
concentration is expressed as a mean recovery percentage.
Sample Dilution factor Expected Measured Mean recovery
concentration (ng/ml) concentration (ng/ml) percentage
PS 51 1/1 127.71 127.71 100
1/2 63.86 66.88 105
1/4 31.93 34.16 107
1/8 15.96 16.32 102
1/16 7.98 8.95 112
1/32 3.99 4.17 104
PS 54 1/1 150.27 150.27 100
1/2 75.14 93.24 124
1/4 37.57 43.35 115
1/8 18.78 22.07 117
1/16 9.39 10.96 117
1/32 4.70 5.70 121
PS 55 1/1 196.57 196.57 100
1/2 98.29 108.00 110
1/4 49.14 52.68 107
1/8 24.57 26.09 106
1/16 12.29 13.29 108
1/32 6.14 7.05 115

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Comparison with another test method INDEX OF SYMBOLS

Correlation was established between the VIDAS CK-MB Symbol Meaning
kit and another commercially available kit (X) using an
enzyme immunoassay technique:
Catalog number
The allometric curve equation obtained is:
VIDAS CK-MB = 1.02 X + 0.33 r = 0.99 (n = 94)
In Vitro Diagnostic Medical Device
WASTE DISPOSAL
Dispose of used or unused reagents as well as any other Manufacturer
contaminated disposable materials following procedures
for infectious or potentially infectious products.
It is the responsibility of each laboratory to handle waste Temperature limit
and effluents produced according to their nature and
degree of hazardousness and to treat and dispose of
them (or have them treated and disposed of) in Use by date
accordance with any applicable regulations.
Batch code
LITERATURE REFERENCES
1. LANDT Y., VAIDYA H.C., PORTER S.E., DIETZLER D.N., Consult Instructions for Use
LADENSON J.H. Immunoaffinity purification of creatine
kinase-MB from human, dog and rabbit heart with use of a Contains sufficient for <n> tests
monoclonal antibody specific for CK-MB. Clin. Chem., 1989,
35 (6), 985-989.
2. REVENKO I., MANCHON M. Intérêt actuel de la CK-MB Date of manufacture
dans le diagnostic de l'infarctus du myocarde. Lyon
pharmaceutique. Elsevier, Paris. 1993, 44 (2), 83-90.
3. GERHARDT W., LJUNGDAHL L. Rational diagnostic WARRANTY
strategy in diagnosis of ischemic myocardial injury. S- bioMérieux disclaims all warranties, express or implied,
troponin T and S-CK-MB (mass) time series using individual
including any implied warranties of MERCHANTABILITY
baselines values. Scand. J. Clin. Lab. Invest., 1993, 53 (215),
47-59. AND FITNESS FOR A PARTICULAR USE. bioMérieux
shall not be liable for any incidental or consequential
4. BHAYANA V., COHOE S., LEUNG F.Y., JABLONSKY G.,
HENDERSON A.R. Diagnostic evaluation of creatine kinase-
damages. IN NO EVENT SHALL BIOMERIEUX’S
2 mass and creatine kinase-3 and -2 isoform ratios in early LIABLITY TO CUSTOMER UNDER ANY CLAIM
diagnosis of acute myocardial infarction. Clin. Chem., 1993, EXCEED A REFUND OF THE AMOUNT PAID TO
39 (3), 488-495. BIOMERIEUX FOR THE PRODUCT OR SERVICE
5. BEN AYED S., BOUKEZIA N., FOGLIETTI MJ.,BERNARD WHICH IS THE SUBJECT OF THE CLAIM.
M. Evaluation de la technique de mesure de la CK-MB par
immuno-enzymofluorimétrie sur VIDAS (bioMérieux). Ann.
Biol. Clin., 1996, 54, 145-149.
6. VAIDYA H.C., BEATTY B.G. Eliminating interference from
heterophilic antibodies in a two-site immunoassay for
creatine kinase MB by using F(ab')2 conjugate and polyclonal
mouse IgG. Clin. Chem., 1992, 38 (9), 1737-1742.
7. VAIDYA H.C., MAYNARD Y., DIETZLER D.N., LADENSON
J.H. Direct measurement of creatine kinase-MB activity in
serum after extraction with a monoclonal antibody specific to
the MB isoenzyme. Clin. chem., 1986, 32 (4), 657-663.
8. LAMPE, A.S., H.J. PEITERSE-BRUINS, and J.C.R.
EGTERVAN WISSERDERKE, 1988, Wearing gloves as
cause of false negative HIV tests. Lancet II, 1140-1144.

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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 07709J 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.
Any other name or trademark is the property of its respective owner.

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