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www.biomaghreb.com
IVD
CREATINE KINASE LS
CK-Nac activeted, IFCC method
FT En 09 Date of updating : 10/ 2020 Version B
IN VITRO DIAGNOSTIC USE
Manufacturer Use by In Vitro Diagnostic Temperature
Limitation
Catalogue number See insert Store away from light Sufcient
for < n > essais
Batch number
CINICAL SIGNIFICANCE
Creatine kinase (CK) is an enzyme which consists of isoenzymes mainly of the muscle (CK-M) and
the brain (CK-B). CK exists in serum in dimeric form as CK-MM, CK-MB, CK-BB and as macro en-
zyme. Elevated CK values are observed in cardiac muscle damages and in skeletal muscle diseases.
Measurement of CK is used especially in conjunction with CK-MB for diagnosis and monitoring of
myocardial infarction.
PRINCIPLE
Kinetic determination of creatine kinase reactivated by N-acetylcysteine according to the following
reactions:
Creatine phosphate + ADP
CK
Creatine + ATP
D- Glucose + ATP
HK
D-Glucose- 6 P + ADP
D-Glucose-6-phosphate- NADP
G-6PDH
D-Gluconate-6-P+ NADPH+H
CK = creatine kinase
HK= Hexokinase
G-6-PDH=Glucose-6-phosphate dehydrogenase. The catalytic activity of CK is determined by mea-
suring the rate of appearance of NADPH+H+ at 340 nm.
REAGENT COMPOSITION
Reagent 1
Buffer imidazole Acetate pH: 6.7
Glucose
100mmol/l
20 mmol/l
Reagent 2
Substrate
N-Acetyl cysteine
Creatine phosphate
ADP
AMP
NADP
Diadenosine pentaphosphate
Hexokinase
Glucose-6-phosphate dehydrogenase
20 mmol/l
30 mmol/l
5 mmol/l
5 mmol/l
2 mmol/l
10 µmol/l
2500 Ul/l
1500 Ul/l
REAGENT PREPARATION
Collect sample using standard sampling tubes.
heparinazed, or EDTA plasma
Stability: 7 days at +4 °C to 8°C
2 days at +20°C to 25°C
Centrifuge samples containing precipitate before performing assay.
PREPARATION AND STABILITY OF THE WORKING SOLUTION
1) Serum start :
Mix 4 volumes of R1 with 1 volume of R2,
Stability : 10 days at 2 - 8°C
1 day at 20 - 25°C
Unopened kit components, and at 2 - 8°C: Up to the expiry date.
2) Substrate (R2) start:
R1: ready to use
R2: ready to use
Onboard stability: R1 21 days.
R2 21 days.
SAFETY CAUTIONS
Biomaghreb reagents are intended for use by qualified personnel for in vitro use (do not pipette by
mouth).
• Refer to the current SDS available on request or at www.biomaghreb.com;
• Verify the integrity of the reagents before use; and
• Disposal of waste: comply with the legislation in force.
For safety reasons, treat any specimen or reagent of biological origin as potentially infectious.
Respect the legislation in force
ADDITIONAL EQUIPMENT
- Basic equipment of the medical analysis laboratory;
- Spectrophotometer or Clinical Biochemistry Analyzer
LINEARITY
Linearity up to 900 U/l at 37° C.
If the ΔDO/min is greater than 0,200 (340 or 334 nm) repeat the test using a sample diluted 1:10 with
saline solution. Multiply result by 10.
PROCEDURE
1) Manual procedure : Serum start
Wavelength 340nm. Hg 334 or Hg 365 nm
Temperature +25 / +30 / +37°C
cuvette 1 cm light path
Zero adjustment air or distilled water
Macro Semi-Micro Micro
Working Solution
2500µl 1000µl 500µl
Sample
100µl 40µl 20µl
Mix and incubate for 2 minutes, measure the absorbance increase per minutes for 3 min
Calculation
Macro Semi-Micro Micro
340nm ΔA/min.x
4130 4130 4130
Hg 334nm ΔA/min.x
4207 4207 4207
Hg 365nm ΔA/min.x
7429 7429 7429
2) Manual procedure : Substrate (R2) start
Wavelength 340nm. Hg 334 or Hg 365 nm
Temperature +25 / +30 / +37°C
cuvette 1 cm light path
Zero adjustment air or distilled water
Macro Semi-Micro Micro
R1
2000µl 800µl 400µl
Sample
100µl 40µl 20µl
R2
500µl 200µl 100µl
Mix and incubate for 2 minutes, measure the absorbance increase per minutes for 3 min.
Calculation
Macro Semi-Micro Micro
340nm ΔA/min.x
4130 4130 4130
Hg 334nm ΔA/min.x
4207 4207 4207
Hg 365nm ΔA/min.x
7429 7429 7429
REFERENCE VALUES
In all cases, each laboratory should establish its own reference values.
T °C
25°C 30°C 37°C
Men
10-80 U/l 15-130 U/l 25-195 U/l
Women
10-70 U/l 15-110 U/l 25-170 U/l
REFERENCES
- Bablok W. et al. A genéral Regression Procedure for Method transformation;
- JClin Chem Clin Biochem 1988 ; 26 : 78-790
- Black H.R. Quallich H gareleck CB. Racial diffrences in serum Creatine kinase levels Am J Med 7986
; 81 : 479-487;
- GLick M.R, Ryder KW, Jackson SA. Graphital Comparisons of Interferences in Clinical Chemistry
Instrumentation Clin Chem 1986 ; 32 ; 470-474 Passing;
- H. Bablok W. A New Biometrical Procedure for Testing the Equality of Measurements from Two
Different Analytical Methods J Clin Chem Clin Biochem 1983; 21: 707-720 ;
- Guder W. G., Narayanan S., Wisser H., Zawta B. List of Analytes Preanalytical Variables, Brochure in
Samples : From The Patient to the Laboratory Darnstadt GIT Veriag 1996.
