CREATINE KINASE BR

REF 1120010 REF 1120005

1 x 25 mL 2 x 50 mL
CREATINE KINASE BR
CK NAC-ACTIVATED
CONTENTS CONTENTS UV enzymatic method
R1. Reagent 1 x 20 mL R1. Reagent 4 x 20 mL
R2. Reagent 1 x 5 mL R2. Reagent 1 x 20 mL
KINETIC

For in vitro diagnostic use only

PRINCIPLE SAMPLES

Creatine kinase (CK) catalyzes the reaction between creatine Serum. Stable 8 days at 2-8ºC or 1 month at –20ºC. Chill the
phosphate (CP) and adenosine 5´-diphosphate (ADP) with samples as rapidily as possible after collection.
formation of creatine and adenosine 5´-triphosphate (ATP). The Moderately or severely hemolyzed specimens are unsatisfactory
latter phosphorylates glucose to glucose-6-phosphate (G6P) in the for testing, as well as plasmas containing EDTA, heparin, citrate, or
presence of hexoquinase (HK). G6P is oxidized to Gluconate-6P in fluoride since they may produce unpredictable reaction rates.
the presence of reduced nicotinamide-adenine dinucleotide
phosphate (NADP) in a reaction catalyzed by glucose-6-phosphate
dehydrogenase (G6P-DH). INTERFERENCES
The conversion is monitored kinetically at 340 nm by the rate of
increase in absorbance resulting from the reduction of NADP to  Lipemia (intralipid 5 g/L) may affect the results.
NADPH proportional to the activity of CK present in the sample.
In this test1,2 the presence of N-acetilcysteine (NAC) allows the  Bilirubin (< 20 mg/dL), hemoglobin (< 10 g/L), do not interfere.
optimal activation of the enzyme.  Other drugs and substances may interfere3.

CK (AMP, NAC)
CP + ADP pH 6.5 Creatine + ATP
MATERIALS REQUIRED
HK
ATP + Glucose ADP + G6P  Photometer or spectrophotometer with a thermostatted cell
G6P-DH compartment set at 25/30/37ºC, capable of reading at 340 nm.
 Stopwatch, strip-chart recorder or printer.
G6P + NADP+ + H2O Gluconate-6P + NADPH + H+
 Cuvettes with 1-cm pathlength.
This test has been formulated according the standarized method  Pipettes to measure reagent and samples.
described by IFCC. Clin Chem Lab Med 2002; 40(6) : 635-642.

PROCEDURE
REAGENT COMPOSITION

R1 Buffer/Glucose/NAC. Imidazol buffer 100 mmol/L pH 6.7, 1. Preincubate working reagent, samples and controls to reaction
glucose 20 mmol/L, NAC 20 mmol/L, magnesium acetate temperature.
10 mmol/L, NADP 2.5 mmol/L, HK  4 KU/L, EDTA 2 2. Set the photometer to 0 absorbance with distilled water.
mmol/L. 3. Pipette into a cuvette:
R2 Substrate/Coenzymes. CP 30 mmol/L, AMP 5 mmol/L, ADP
2 mmol/L, di(adenosine-5´) pentaphosphate 10 mol/L, G6P- Reaction temperature 25ºC 30ºC 37ºC
DH  1.5 KU/L.
Working reagent 1.0 mL 1.0 mL 1.0 mL
STORAGE AND STABILITY Sample or control 40 L 40 L 20 L

Store at 2-8ºC.
All the kit compounds are stable until the expiry date stated on the 4. Mix gently by inversion. Insert cuvette into the cell holder and
label. Do not use reagents over the expiration date. start stopwatch.
Store the vials tightly closed, protected from light and prevented 5. Incubate for 3 minutes and record initial absorbance reading.
contaminations during the use. 6. Repeat the absorbance readings exactly after 1, 2 and 3
Discard If appear signs of deterioration: minutes.
- Presence of particles and turbidity. 7. Calculate the difference between absorbances.
- Blank absorbance (A) at 340 nm > 0.400 in 1cm cuvette. 8. Calculate the mean of the results to obtain the average change
in absorbance per minute (A/min).

REAGENT PREPARATION
CALCULATIONS
Working reagent. Mix 4 mL of R1 + 1 mL of R2. Stable for 1 month at
2-8ºC or 10 days at 16-25ºC. A/min x 4127 = U/L CK (25/30ºC)
Protect from light. A/min x 8095 = U/L CK (37ºC)

QUALITY SYSTEM CERTIFIED LINEAR CHEMICALS, S.L.U. Joaquim Costa 18 2ª planta. 08390 Montgat (Barcelona) SPAIN
ISO 9001 ISO 13485 Telf. (+34) 934 694 990; E-mail: [email protected] ; website: www.linear.es NIF-VAT:B60485687
Samples with A/min exceeding 0.270 a 340 nm should be diluted ANALYTICAL PERFORMANCE
1:10 with saline and assayed again. Multiply the results by 10.
- Detection Limit : 10.11 U/L
If results are to be expressed as SI units apply:
U/L x 16.67 = nkat/L - Linearity : Up to 1000 U/L
- Precision:
U/L Within-run Between-run
REFERENCE VALUES1 Mean 227 564 227 564
Serum SD 2.62 13.12 1.15 8.13
CV% 1.15 2.30 2.52 1.44
Temperature 25ºC 30ºC 37ºC N 10 10 10 10
 65 U/L  105 U/L  174 U/L - Sensitivity : 0.125 mA / min/ U/L CK-NAC.
Men
(1083 nkat/L) (1750 nkat/L) (2900 nkat/L)
- Correlation: This assay (y) was compared with a similar commercial
 55 U/L  80 U/L  140 U/L method (x). The results were:
Women
(917 nkat/L) (1334 nkat/L) (2334 nkat/L) N = 25 r = 0.999 y = 1.098x + 6.8
 94 U/L  150 U/L  225 U/L The analytical performances have been generated using on
Children
(1570 nkat/L) (2500 nkat/L) (3750 nkat/L) automatic instrument. Results may vary depending on the
instrument.
It is recommended that each laboratory establishes its own
reference range.
NOTES

QUALITY CONTROL 1. This method may be used with different instruments. Any
application to an instrument should be validated to demonstrate
To ensure adequate quality control (QC), each run should include a that results meets the performance characteristics of the
set of controls (normal and abnormal) with assayed values handled method. It is recommended to validate periodically the
as unknowns. instrument. Contact to the distributor for any question on the
application method.
REF 1980005 HUMAN MULTISERA NORMAL 2. Clinical diagnosis should not be made on findings of a single test
Borderline level of CK. Assayed. result, but should integrate both clinical and laboratory data.

REF 1985005 HUMAN MULTISERA ABNORMAL

Elevated level of CK. Assayed.
REFERENCES
If the values are found outside of the defined range, check the
instrument, reagents and procedure. 1. Szasz, G., Grober, W and Bernt, E. Clin. Chem. 22 : 650 (1976).
Each laboratory should establish its own Quality Control scheme 2. German Society for Clinical Chemistry: Recommendations of the
and corrective actions if controls do not meet the acceptable Enzyme Commision. J. Clin. Chem. Clin. Biochem. 15 : 255 (1977).
tolerances. 3. Young DS. Effects of drugs on clinical laboratory tests, 5th ed.
AACC Press, 2000.
4. Auvinen, S. Acta. Med. Scand. (Suppl. 539) (1972).
CLINICAL SIGNIFICANCE 5. Doran, G.R., and Wilkinson, J.H. Clin. Chim. Acta. 62 : 203
(1975).
Creatine kinase (CK) values are high in patients with myocardial 6. Fisher, M.D., Carliner, N.H., Becker, L.C., Peters, R.W. y
infarction, progressive muscular dystrophy, alcoholic myopathy, and Plotnick, G.D. J.A.M.A. 249 : 393 (1983).
delirium tremens, but normal in patients with hepatitis and other
forms of liver disease. The high values in patients with
hypothyroidism reflect the muscle changes in this condition.
Although CK is found almost exclusively in myocardium, muscle,
and brain and early reports suggested it to be an almost specific
index of injury of myocardium and muscle, more recent reports
indicate that, inexplicably high serum CK values can occur in
patients with pulmonary infarction and pulmonary edema.
At present, it should be regarded as a useful but not completely
specific adjunt in the diagnosis of myocardial and muscle disease.
Specificity of CK assay is enhanced by measurement of its
isoenzymes.4-6

B1120-2/0901
R1.ing

QUALITY SYSTEM CERTIFIED LINEAR CHEMICALS, S.L.U. Joaquim Costa 18 2ª planta. 08390 Montgat (Barcelona) SPAIN
ISO 9001 ISO 13485 Telf. (+34) 934 694 990; E-mail: [email protected] ; website: www.linear.es NIF-VAT:B60485687