0107190808190COINV1.

CKMB
Creatine Kinase-MB Enzymes
Order information
Analyzer(s) on which cobasc pack(s) can be used
07190808 190 Creatine KinaseMB (100tests) SystemID0774847 COBASINTEGRA 400 plus
11447394 216 Calibrator f.a.s. CKMB (3 x 1mL) System-ID 0779962
11447378 122 Precinorm CKMB (4 x 3mL) System-ID 0791113
04358210 190 Precipath CK-MB (4 x 3mL) System-ID 0768286
05117003 190 PreciControl ClinChem Multi 1 (20 x 5mL) System-ID 0774693
05947626 190 PreciControl ClinChem Multi 1 (4 x 5mL) System-ID 0774693
05117216 190 PreciControl ClinChem Multi 2 (20 x 5mL) System-ID 0774707
05947774 190 PreciControl ClinChem Multi 2 (4 x 5mL) System-ID 0774707
20756350 322 NaCl Diluent 9 % (6 x 22 mL) System-ID 0756350

English SR CAPSO* buffer: 20mmol/L, pH8.8 (37C); glucose: 120mmol/L;

System information EDTA: 2.46mmol/L; creatine phosphate: 184mmol/L;
Test CKMB2, test ID 0-057 4monoclonal antiCKM antibodies (mouse), inhibitingcapacity:
>99.6%up to 66.8kat/L(4000U/L) (37C) CKM subunit;
Intended use preservative; stabilizers; additive.
In vitro test for the quantitative determination of the catalytic activity of
creatine kinaseMB subunit (CKMB) in human serum and plasma on *CAPSO: 3-(cyclohexylamino)-2-hydroxy-1-propanesulfonic acid
COBASINTEGRA systems. R1 is in position B and SR is in position C.
Summary Precautions and warnings
Creatine kinase(CK) appears as three isoenzymes which are dimers Pay attention to all precautions and warnings listed in
composed of two types of monomer subunits. The isoenzymes comprise all Section1/Introduction of this Method Manual.
three combinations of monomers, M(for skeletal muscle derived) and B(for
brain derived), as represented by the notations MM, MB, and BB.1 This kit contains components classified as follows in accordance with the
Regulation (EC) No.1272/2008:
Many organs contain CK, but the distribution of isoenzymes is different in
each one. Skeletal muscle is very rich in the MM isoenzyme, while brain,
stomach, intestine, bladder, and lung contain primarily the BB isoenzyme.
The MB isoenzyme has been found in appreciable amounts (15 to
20percent) only in myocardial tissue. Therefore, total serum CK activity is
elevated in a number of diseases. This lack of specificity limits its diagnostic
value. However, the striking difference in the CK isoenzyme patterns from Danger
different organs has made CK one of the most useful enzymes for
diagnostic purposes in acute myocardial infarction. CKMB appears in
serum reflecting its unique presence in myocardial tissue. It is in supporting H360D May damage the unborn child.
the diagnosis of suspected myocardial infarction that serial determinations Prevention:
of CK isoenzymes find their most frequent application in the clinical
laboratory.1,2 P201 Obtain special instructions before use.
After immunoinhibition with antibodies to the CKM subunit,3 the CKB
activity is determined with a standardized method for the determination of P202 Do not handle until all safety precautions have been read
CK with activation by NAC as recommended by the German Society for and understood.
Clinical Chemistry (DGKC)4 and the International Federation of Clinical
Chemistry (IFCC)5,6 in 1977 and 2002 respectively. This assay meets the P280 Wear protective gloves/ protective clothing/ eye protection/
recommendations of the IFCC and DGKC, but was optimized for face protection.
performance and stability.
Response:
Test principle
Immunological UV assay P308 + P313 IF exposed or concerned: Get medical advice/attention.
Sample and addition of R1 (buffer/enzymes/coenzyme) Storage:
Addition of R2 (buffer/substrate/antibody) and start of reaction.
P405 Store locked up.
Human CKMB is composed of two subunits, CKM and CKB which both
have an active site. With the aid of specific antibodies to CKM, the catalytic Disposal:
activity of CKM subunits in the sample is inhibited to 99.6% without
affecting the CKB subunits. The remaining CKB activity, corresponding to P501 Dispose of contents/container to an approved waste
half the CKMB activity, is determined by the total CK method. As the disposal plant.
CKBB isoenzyme only rarely appears in serum and the catalytic activity of Product safety labeling primarily follows EU GHS guidance.
the CKM and CKB subunits hardly differ, the catalytic activity of the
CKMB isoenzyme can be calculated from the measured CKB activity by Contact phone: all countries: +49-621-7590
multiplying the result by 2. Reagent handling
Reagents - working solutions Ready for use
R1 Imidazole buffer: 123mmol/L, pH6.5 (37C); EDTA: 2.46mmol/L; Storage and stability
Mg2+: 12.3mmol/L; ADP: 2.46mmol/L; AMP: 6.14mmol/L; Shelf life at 28C See expiration date on
diadenosine pentaphosphate: 19mol/L; NADP (yeast): cobasc pack label
2.46mmol/L; Nacetylcysteine: 24.6mmol/L; HK (yeast):
36.7kat/L; G6PDH (E.coli):23.4kat/L; preservative; Onboard in use at 1015C 8weeks
stabilizers; additives.

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CKMB
Creatine Kinase-MB Enzymes

Specimen collection and preparation Calibration

For specimen collection and preparation only use suitable tubes or
collection containers. Calibrator C.f.a.s. CKMB
Only the specimens listed below were tested and found acceptable. Use deionized water as zero
Serum: Nonhemolyzed serum is the specimen of choice and also calibrator.
recommended by IFCC.
Plasma: Liheparin, K2, K3EDTA plasma. Calibration mode Linear regression
Liheparin in the usual concentration does not interfere with the test, but Calibration replicate Duplicate recommended
IFCC warns against its use.5 Calibration interval Each lot and as required following
The sample types listed were tested with a selection of sample collection quality control procedures
tubes that were commercially available at the time of testing, i.e. not all
available tubes of all manufacturers were tested. Sample collection systems Traceability: This method has been standardized against the IFCC Method
from various manufacturers may contain differing materials which could for Creatine Kinase6 with addition of antibodies.
affect the test results in some cases. When processing samples in primary Quality control
tubes (sample collection systems), follow the instructions of the tube
manufacturer. Reference range Precinorm CKMB or
Centrifuge samples containing precipitates before performing the assay. PreciControlClinChemMulti1
Stability in serum:7 8hours at 2024C Pathological range Precipath CKMB or
PreciControlClinChemMulti2
8days at 28C
Control interval 24hours recommended
4weeks at -20C
Control sequence User defined
Stability in heparin plasma:7 8hours at 2024C
Control after calibration Recommended
5days at 28C
For quality control, use control materials as listed in the Order information
8days at -20C section. In addition, other suitable control material can be used.
Stability in EDTA plasma:8 2days at 2025C The control intervals and limits should be adapted to each laboratorys
7days at 48C individual requirements. Values obtained should fall within the defined
limits. Each laboratory should establish corrective measures to be taken if
1year at -20C values fall outside the defined limits.
Materials provided Follow the applicable government regulations and local guidelines for
quality control.
See Reagents working solutions section for reagents.
Calculation
Materials required (but not provided)
COBASINTEGRAanalyzers automatically calculate the analyte activity of
NaClDiluent9%, Cat.No.20756350322, systemID0756350 for each sample. For more details, please refer to Data Analysis in the Online
automatic sample dilution. NaClDiluent9% is placed in its predefined rack Help (COBASINTEGRA400plus analyzer).
position and is stable for 4weeks onboard COBASINTEGRA400plus
analyzers. Conversion factor: U/Lx0.0167=kat/L
Assay Limitations - interference
For optimum performance of the assay follow the directions given in this The total CK activity of the specimen should be determined prior to
document for the analyzer concerned. Refer to the appropriate operators performing the CKMB assay. The amount of antihuman CKM subunit
manual for analyzerspecific assay instructions. antibody in the CKMB reagent is sufficient for the complete inhibition of up
Application for serum and plasma to 4000U/L CKM activity. If the total CK activity exceeds 4000U/L, the
specimen requires dilution because complete inhibition of the CKM subunit
COBASINTEGRA400plus test definition is no longer assured. In patients with a disposition to macroCK formation,
Measuring mode Absorbance implausibly high CKMB values may be measured in relation to the total CK,
since the macroforms mainly consist of CKB subunits. As these patients
Abs. calculation mode Kinetic have generally not suffered a myocardial infarction, additional diagnostic
measures are necessary.9
Reaction mode R1SSR
Criterion: Recovery within 10% of initial value at a creatine kinaseMB
Reaction direction Increase activity of 25U/L (0.42kat/L).
Wavelength A/B 340/552nm Icterus:10 No significant interference up to an Iindex of 60 for conjugated
and 20 for unconjugated bilirubin (approximate conjugated bilirubin
Calc. first/last 10/5069 concentration: 1026mol/L or 60mg/dL and approximate unconjugated
Unit U/L bilirubin concentration: 342mol/L or 20mg/dL).
Hemolysis:10 No significant interference up to an Hindex of 20
Pipetting parameters (approximate hemoglobin concentration: 12.4mol/L or 20mg/dL).
Diluent (H2O) Lipemia (Intralipid):10 No significant interference up to an Lindex of 500.
There is poor correlation between the Lindex (corresponds to turbidity) and
R1 100L - triglycerides concentration. Choose diluted sample treatment for automatic
Sample 5L - rerun.
SR 20L - Adenylate kinase: Adenylate kinase (AK) may cause positive interference.
Sources of AK in the blood are erythrocytes, muscle, and liver. In order to
Total volume 125L reduce AK interference to a minimum, AMP and Ap5A are included in the
reagent. The AMP/Ap5A mixture causes 97% inhibition of the AK from
erythrocytes and muscle, and 95% inhibition of the AK from liver.4 The
slight residual AK activity does not influence the assay of total CK, but may
affect the low CKMB activities.

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CKMB
Creatine Kinase-MB Enzymes

Drugs: No interference was found at therapeutic concentrations using activity is used, the diagnostic efficiency will be lower and will vary with the
common drug panels.11,12 sampling time.1,9
Exceptions: Cyanokit (hydroxocobalamin), Cefoxitin, Sulfasalazine and Each laboratory should investigate the transferability of the expected values
Sulfapyridine at therapeutic concentrations interfere with the test. to its own patient population and if necessary determine its own reference
In very rare cases, gammopathy, in particular type IgM (Waldenstrms ranges.
macroglobulinemia), may cause unreliable results.13 Specific performance data
For diagnostic purposes, the results should always be assessed in Representative performance data on the COBASINTEGRA analyzers are
conjunction with the patients medical history, clinical examination and other given below. Results obtained in individual laboratories may differ.
findings.
Precision
ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory Repeatability and intermediate precision were determined using human
when certain test combinations are run together on COBASINTEGRA samples and controls in accordance with the CLSI (Clinical and Laboratory
analyzers. Refer to the CLEAN Method Sheet for further instructions and for Standards Institute) EP5 requirements (2aliquots per run, 2runs per day,
the latest version of the Extra wash cycle list. 21days). The following results were obtained:
Where required, special wash/carry-over evasion programming must Repeatability Mean SD CV
be implemented prior to reporting results with this test.
Limits and ranges U/L (kat/L) U/L (kat/L) %
Measuring range Human serum 1 22.2 (0.37) 0.7 (0.01) 3.2
32000U/L (0.0533.4kat/L)
Human serum 2 31.6 (0.53) 0.5 (0.01) 1.7
Determine samples having higher activities via the rerun function. Dilution
of samples via the rerun function is a 1:3 dilution. Results from samples Human serum 3 562 (9.39) 3.0 (0.05) 0.5
diluted by the rerun function are automatically multiplied by a factor of 3. Human serum 4 1105 (18.5) 7.0 (0.12) 0.6
Lower limits of measurement
Human serum 5 1949 (32.6) 27 (0.45) 1.4
Limit of Blank, Limit of Detection and Limit of Quantitation PCCC Multi 1* 44.5 (0.74) 0.6 (0.01) 1.3
Limit of Blank = 3 U/L (0.05 kat/L) PCCC Multi 2 106 (1.77) 0.8 (0.01) 0.7
Limit of Detection = 3 U/L (0.05 kat/L)
Intermediate precision Mean SD CV
Limit of Quantitation = 5 U/L (0.08 kat/L)
The Limit of Blank, Limit of Detection and Limit of Quantitation were U/L (kat/L) U/L (kat/L) %
determined in accordance with the CLSI (Clinical and Laboratory Standards Human serum 1 22.2 (0.37) 0.8 (0.01) 3.8
Institute) EP17A2 requirements.
The Limit of Blank is the 95th percentile value from n60 measurements of Human serum 2 31.6 (0.53) 0.7 (0.01) 2.2
analytefree samples over several independent series. The Limit of Blank Human serum 3 562 (9.39) 5.0 (0.08) 0.9
corresponds to the concentration below which analytefree samples are
found with a probability of 95%. Human serum 4 1085 (18.1) 9.8 (0.16) 0.9
The Limit of Detection is determined based on the Limit of Blank and the Human serum 5 1949 (32.6) 34 (0.57) 1.7
standard deviation of low concentration samples. The Limit of Detection
corresponds to the lowest analyte concentration which can be detected PCCC Multi 1 43.5 (0.73) 0.8 (0.01) 1.8
(value above the limit of blank with a probability of 95%). PCCC Multi 2 104 (1.74) 1.6 (0.03) 1.5
The Limit of Quantitation is the lowest analyte concentration that can be *PCCC=PreciControl ClinChem
reproducibly measured with a precision of 20%CV. It has been determined
using low concentration creatine kinaseMB samples. Method comparison
Expected values Creatine kinaseMB values for human serum and plasma samples obtained
Reference intervals strongly depend on the patient group regarded and the on a COBASINTEGRA400plus analyzer(y) were compared with those
specific clinical situation. using the corresponding reagent on a Roche/Hitachi MODULARP
analyzer(x).
For healthy people: Reference range (37C) according toKleinetal.14 and
consensus values:15 Sample size (n)=117

<25U/L (<0.418kat/L) Passing/Bablok18 Linear regression

For myocardial infarction diagnosis using the combination CK and CKMB y=1.016x+3.75U/L y=1.011x+4.26U/L
(activity), and representing a CK consensus value based on longterm =0.897 r=1.000
experience:15,16
The sample activities were between 5.0 and 1967U/L (0.08 and
1. CKmen >190U/L (3.17kat/L) 32.8kat/L).
CKwomen >167U/L (2.79kat/L) References
2. CKMB >24U/L (0.40kat/L) 1 Moss DW, Henderson AR, Kachmar JF. Enzymes. In: Tietz NW, ed.
Fundamentals of Clinical Chemistry, 3rd ed. Philadelphia, PA: WB
3. The CKMB activity accounts for 625% of the total CK activity. Saunders 1987;346-421.
When myocardial infarction is suspected the diagnostic strategy proposals 2 Lott JA, Stang JM. Serum enzymes and isoenzymes in the diagnosis
in the consensus document of European and American cardiologists should and differential diagnosis of myocardial ischemia and necrosis. Clin
in general be followed.17 Chem 1980;26:1241-1250.
If despite the suspicion of myocardial infarction the values found remain 3 Wrzburg U, Hennrich N, Lang H, et al. Determination of creatine
below the stated limits, a fresh infarction may be involved. In such cases kinase-MB in serum using inhibiting antibodies. Klin Wschr
the determinations should be repeated after 4hours. 1976;54(8):357-360.
Maximum diagnostic efficiency of the CKMB determination will be obtained
when a sequential sampling protocol is used and consideration is given to
the time pattern of activity over a 6 to 48hour period. When only CKMB

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CKMB
Creatine Kinase-MB Enzymes

4 Bergmeyer HU, Breuer H, Bttner H, et al. Empfehlungen der

Deutschen Gesellschaft fr Klinische Chemie. Standard-Methode zur
Bestimmung der Aktivitt der Creatin-Kinase. J Clin Chem Clin Roche Diagnostics GmbH, SandhoferStrasse116, D-68305 Mannheim
Biochem 1977;15:249-254. www.roche.com
5 Hrder M, Elser RC, Gerhardt W, et al. IFCC methods for the
measurement of catalytic concentration of enzymes. Provisional
recommendation IFCC method for creatine kinase Appendix A. J Int
Fed Clin Chem 1990;2:26-35.
6 Schumann G, Bonora R, Ceriotti F, et al. IFCC Primary Reference
Procedures for the Measurement of Catalytic Activity Concentrations of
Enzymes at 37 C Part 2. Reference Procedure for the Measurement
of Catalytic Concentration of Creatine Kinase. Clin Chem Lab Med
2002;40(6):635-642.
7 Braun S, Rschenthaler F, Jarausch J, et al. Analyte Stability of CK-MB
Activity and cTnT in ICU Patient Serum and Heparin Plasma. Poster
presented at Medica 2004, Dsseldorf. (Roche Diagnostics GmbH No.
04587979990).
8 Use of Anticoagulants in Diagnostic Laboratory Investigations. WHO
Publication WHO/DIL/LAB/99.1 Rev. 2. Jan. 2002.
9 Remaley AT, Wilding P. Macroenzymes: Biochemical Characterization,
Clinical Significance, and Laboratory Detection. Clin Chem
1989;35:2261-2270.
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 Bakker AJ, Mcke M. Gammopathy interference in clinical chemistry
assays: mechanisms, detection and prevention.
ClinChemLabMed2007;45(9):1240-1243.
14 Klein G, Berger A, Bertholf R, et al. Abstract: Multicenter Evaluation of
Liquid Reagents for CK, CK-MB and LDH with Determination of
Reference Intervals on Hitachi Systems. Clin Chem 2001;47:Suppl.
A30.
15 Thomas L, Mller M, Schumann G, et al. Consensus of DGKL and
VDGH for interim reference intervals on enzymes in serum. J Lab Med
2005; 29(5):301-308.
16 Stein W. Strategie der klinischen-chemischen Diagnostik des frischen
Myokardinfarktes. Med Welt 1985;36:572-577.
17 Myocardial Infarction Redefined - A Consensus Document of the Joint
European Society of Cardiology/ American College of Cardiology
Committee for the Redefinition of Myocardial Infarction. Eur Heart J
2000;21:1502-1513.
18 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.
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.
Symbols
Roche Diagnostics uses the following symbols and signs in addition to
those listed in the ISO 152231 standard:
Contents of kit
Volume after reconstitution or mixing
GTIN Global Trade Item Number

COBAS, COBASC, PRECICONTROL, PRECINORM and PRECIPATH are trademarks of Roche.

All other product names and trademarks are the property of their respective owners.
Additions, deletions or changes are indicated by a change bar in the margin.
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