CK-MB liquiUV Pipette into cuvettes 37°C

Sample / [CAL] 50 μl
Liquid NAC activated UV Test
[ENZ] 1000 μl
Creatine Kinase (EC 2.7.3.2) Mix, incubate 3 minutes at the desired temperature.
Package Size [SUB] 250 μl
[REF] 12118 10 x 10 ml Complete test kit
Mix and incubate for 3 minutes. Read the absorbance and at the same
[IVD]
time start the stopwatch. Read the absorbance again exactly after 1, 2
Method 1,2 and 3 minutes.
Immunoinhibition method whereby specific antibodies inhibit the activity Calculation
of the CK-M subunit without affecting the activity of the CK-B subunit.
Calculate the mean absorbance change per minute (ΔA/min) and multiply
Because the CK-BB activity in the circulation is negligible, the activity
ΔA/min with the following factors to get the activity in the sample:
measured by this method and multiplied with a factor of 2 reflects the
activity of CK-MB. Sample start Reagent start
Reaction Principle Wavelength 37°C 37°C
CK Hg 334 nm 6796 8414
Creatine phosphate + ADP creatine + ATP 340 nm 6666 8254
HK Hg 365 nm 12000 14857
Glucose + ATP glucose-6-phosphate + ADP Conversion factor of traditional units (U/l) into SI-units (kat/l):
G6P-DH 1 U/l = 16.67 x 10-3 μkat/l
Glucose-6-P + NADP gluconate-6-P + NADPH + H+ 1 μkat/l = 60 U/l
Contents Performance Characteristics
[ENZ] 10 x 8 ml Enzymes Linearity
Imidazole buffer (pH 6.2) 125 mmol/l If the absorbance change per minute (ΔA/min) exceeds
Glucose 25 mmol/l Hg 334 nm/340 nm: ΔA/min = 0.200
Magnesium acetate 12.5 mmol/l Hg 365 nm: ΔA/min = 0.100
EDTA 2.5mmol/l dilute 0.1 ml of the sample with 1.0 ml physiological saline (0.9%) and
AMP 6.25 mmol/l repeat the assay using this dilution. Multiply the results by 11.
N-acetylcysteine 0.25 mmol/l Typical performance data can be found in the Verification Report
Diadenosine pentaphosphate 12.5 μmol/l accessible via
NADP 2.5 mmol/l www.human.de/data/gb/vr/en-ckluv.pdf or
Hexokinase ≥ 5 U/ml www.human-de.com/data/gb/vr/en-ckluv.pdf
SH-stabiliser 31.25 mmol/l
monoclonal-CK antibodies (mouse) Reference Range for Myocardial Infarction (MI)7
blocking capacity up to 2000 U/l CK-MM The likelihood of MI existence is high if the following 3 criteria are met:
Sodium azide 0.095 % Temperature 37°C
[SUB] 2 x 10 ml Substrate 1. Total CK IFCC
ADP 10 mmol/l Men > 190 U/l > 171 U/l
Glucose-6-Phosphate-Dehydrogenase ≥ 14 U/ml Women > 167 U/l > 145 U/l
Creatine phosphate 150 mmol/l
2. CK-MB > 24 U/l > 24 U/l
Sodium azide 0.095 %
3. CK-MB activity ranging between 6% and 25% of the total CK activity.
Additional material but not supplied with the kit
[REF] 13611 Quality Control
[Low/High] 4 x 2 ml CK-MB CONTROL All control sera with CK-MB values determined by this method can be em-
Human serum (lyophilised) ployed. Only control sera with human CK can be used.
[REF] 13612 Automation
[CAL] 2 x 1 ml CK-MB CALIBRATOR Proposals to apply the reagents on analysers are available on request.
Human serum (lyophilised) Each laboratory has to validate the application in its own responsibility.
Reagent Preparation Notes
[ENZ] and [SUB] are ready to use for the substrate start method. 1. Avoid hemolytic samples as erythrocytes may release CK activity which
To prepare the Working reagent, mix 4 parts [ENZ] with 1 part [SUB], e.g. interferes with the test.
8 ml [ENZ] + 2 ml [SUB]. 2. Lipaemia: Intralipid shows no effect up to 1000 mg/dl, though
Reagent Stability triglycerides may interfere at concentrations above 800 mg/dl.
The reagents [ENZ] and [SUB] are stable up to the stated expiry date when 3. Formation of macro-CK, containing mainly CK-B subunits, in some
sealed and stored at 2...8°C. After opening the reagents are stable for patients may lead to implausible high CK-MB values relative to total CK
30 days at 2...8°C. Contamination of the reagents must be avoided. activity. These patients normally have no AMI and need further
The Working reagent is stable for 30 days at 2...8°C and for 2 days at diagnostic clarification.
15...25°C. 4. [ENZ] and [SUB] contain sodium azide (0.095%) as preservative. Do not
swallow. Avoid contact with skin and mucous membranes!
Specimen
Serum, heparinised plasma or EDTA plasma. References
Loss of activity within 7 days at +4°C or within 24 hours at +25°C: 2%. 1. Würzburg U. et al., Klin. Wschr. 54, 357 (1976)
2. Würzburg U. et al., J. Clin. Chem. Clin. Biochem. 15, 131 (1977)
Assay
3. Stein, W., Med. Welt 36, 572-577 (1985)
Wavelength: Hg 365 nm, 340 nm or Hg 334 nm 4. Szasz G., Busch E.W., Abstract presented at 3rd Eur. Congr. Clin. Chem.,
Optical path: 1 cm Brighton/UK, 1979, 3-8
Temperature: 37°C 5. Klauke R. et al., Eur. J. Clin. Chem. Clin. Biochem. 15, 901-909 (1993)
Measurement: against air (increasing absorbance) 6. Horder M., Elser R. et al., Eur. J. Clin. Chem. Clin. Biochem. 29, 435 (1991)
Pipetting Scheme for Sample Start: 7. Thomas L., Labor und Diagnose, TH-Books, 89-97 (2008)
Bring working reagent to the desired temperature and keep the
temperature constant (± 0.5°C) for the duration of the test.
EN-CKMBL INF 1211801 GB 09-2012-13
Pipette into cuvettes 37°C
Sample / [CAL] 50 μl
Working reagent 1000 μl
Mix and incubate at the desired temperature for 5 minutes. Read the
absorbance and at the same time start the stopwatch. Read the
absorbance again exactly after 1, 2 and 3 minutes.
Pipetting Scheme for Reagent Start:
Bring reagents [ENZ] and [SUB] to the desired temperature and keep the Human Gesellschaft für Biochemica und Diagnostica mbH
Max-Planck-Ring 21 · 65205 Wiesbaden · Germany
temperature constant (± 0.5°C) for the duration of the test. Telefon +49 6122-9988-0 · Telefax +49 6122-9988-100 · e-Mail human@human.de