CK-MB liquiUV Pipette into cuvettes 25°C/30°C 37°C

Sample 100 µl 50 µl
Liquid NAC activated UV Test [ENZ] 1000 µl 1000 µl
Creatine Kinase (EC 2.7.3.2) Mix, incubate 3 minutes at the desired temperature.
Package Size [SUB] 250 µl 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 ( A/min) and
Because the CK-BB activity in the circulation is negligible, the activity multiply A/min with the following factors to get the activity in the
measured by this method and multiplied with a factor of 2 reflects the sample:
activity of CK-MB.
Sample start Reagent start
Reaction Principle Wavelength 25°C, 30°C 37°C 25°C, 30°C 37°C
CK
Hg 334 nm 3560 6796 4369 8414
Creatine phosphate + ADP creatine + ATP
340 nm 3492 6666 4286 8254
HK
Glucose + ATP glucose-6-phosphate + ADP Hg 365 nm 6286 12000 7714 14857
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 ( A/min) exceeds
Glucose 25 mmol/l
Magnesium acetate 12.5 mmol/l Hg 334 nm/340 nm: A/min = 0.200
EDTA 2.5mmol/l Hg 365 nm: A/min = 0.100
AMP 6.25 mmol/l dilute 0.1 ml of the sample with 1.0 ml physiological saline (0.9%) and
N-acetylcysteine 0.25 mmol/l repeat the assay using this dilution. Multiply the results by 11.
Diadenosine pentaphosphate 12.5 µmol/l Typical performance data can be found in the Verification Report
NADP 2.5 mmol/l accessible via
Hexokinase ≥ 5 U/ml www.human.de/data/gb/vr/en-ckluv.pdf or
SH-stabiliser 31.25 mmol/l www.human-de.com/data/gb/vr/en-ckluv.pdf
anti-CK antibodies (goat)
Reference Range for Myocardial Infarction (MI)7,8
blocking capacity up to 2000 U/l CK-MM
Sodium azide 0.095 % The likelihood of MI existence is high if the following 3 criteria are met:
[SUB] 2 x 10 ml Substrate Temperature 25°C 30°C 37°C
ADP 10 mmol/l 1. Total CK IFCC
Glucose-6-Phosphate-Dehydrogenase ≥ 14 U/ml Men > 80 U/l > 130 U/l > 195 U/l > 171 U/l
Creatine phosphate 150 mmol/l Women > 70 U/l > 110 U/l > 170 U/l > 145 U/l
Sodium azide 0.095 %
2. CK-MB > 10 U/l > 16 U/l > 25 U/l > 24 U/l
Reagent Preparation
3. CK-MB activity ranging between 6% and 25% of the total CK activity.
[ENZ] and [SUB] are ready to use for the substrate start method.
To prepare the Working reagent, mix 4 parts [ENZ] with 1 part [SUB], e.g. Quality Control
8 ml [ENZ] + 2 ml [SUB]. All control sera with CK-MB values determined by this method can be em-
Reagent Stability ployed. Only control sera with human CK can be used.
The reagents [ENZ] and [SUB] are stable up to the stated expiry date when Automation
sealed and stored at 2...8°C. After opening the reagents are stable for Proposals to apply the reagents on analysers are available on request.
30 days at 2...8°C. Contamination of the reagents must be avoided. Each laboratory has to validate the application in its own responsibility.
The Working reagent is stable for 30 days at 2...8°C and for 2 days at
Notes
15...25°C.
1. Avoid hemolytic samples as erythrocytes may release CK activity which
Specimen interferes with the test.
Serum, heparinised plasma or EDTA plasma. 2. Lipaemia: Intralipid shows no effect up to 1000 mg/dl, though
Loss of activity within 7 days at +4°C or within 24 hours at +25°C: 2%. triglycerides may interfere at concentrations above 800 mg/dl.
Assay 3. Formation of macro-CK, containing mainly CK-B subunits, in some
Wavelength: Hg 365 nm, 340 nm or Hg 334 nm patients may lead to implausible high CK-MB values relative to total CK
Optical path: 1 cm activity. These patients normally have no AMI and need further
Temperature: 25°C, 30°C or 37°C diagnostic clarification.
Measurement: against air (increasing absorbance) 4. [ENZ] and [SUB] contain sodium azide (0.095%) as preservative. Do not
swallow. Avoid contact with skin and mucous membranes!
Pipetting Scheme for Sample Start:
Bring working reagent to the desired temperature and keep the References
temperature constant ( 0.5°C) for the duration of the test. 1. Würzburg U. et al., Klin. Wschr. 54, 357 (1976)
2. Würzburg U. et al., J. Clin. Chem. Clin. Biochem. 15, 131 (1977)
Pipette into cuvettes 25°C, 30°C 37°C
3. Stein, W., Med. Welt 36, 572-577 (1985)
Sample 100 µl 50 µl 4. Szasz G., Busch E.W., Abstract presented at 3rd Eur. Congr. Clin. Chem.,
Working reagent 1000 µl 1000 µl Brighton/UK, 1979, 3-8
Mix and incubate at the desired temperature for 5 minutes. Read the 5. Klauke R. et al., Eur. J. Clin. Chem. Clin. Biochem. 15, 901-909 (1993)
absorbance and at the same time start the stopwatch. Read the 6. Horder M., Elser R. et al., Eur. J. Clin. Chem. Clin. Biochem. 29, 435 (1991)
absorbance again exactly after 1, 2 and 3 minutes. 7. Schumann G. et al., Clin. Chem. .Acta 327, 69-79 (2003)
8. Tietz N. W. (ed.), Clinical Guide to Laboratory Test; 3rd edition, WB Saunders Co,
Pipetting Scheme for Reagent Start: (1995)
Bring reagents [ENZ] and [SUB] to the desired temperature and keep the
temperature constant ( 0.5°C) for the duration of the test. EN-CKMBL INF 1211801 GB 02-2011-11 |

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