[LAB] TOTAL CK, CK-MB Flashcards
REFERENCE METHOD OF TOTAL CK ASSAYS
KINETIC METHOD BY OLIVER-ROSALKI
MODIFIED BY SZASZ
FUNCTION OF CK
CATALYZING THE REVERSIBLE REACTION BETWEEN CREATININE AND ATP TO FORM CREATININE PHOSPHATE AND ADP
CK ACTIVITY IN ORGANSE
HIGHEST TO LOWEST ACTIVITY
SKELETAL MUSCLE, HEART, BRAIN
CLINICAL CONDITIONS CK IS USED TO AID IN DIAGNOSING
MUSCULAR DYSTROPHY
MYOCARDIAL INFARCTION
HYPOTHYROIDISM
RENAL DISEASES
BASIS OF THE EARLY PROCEDURE OF DETERMINING CK
RATE OF ATO FORMATION
SUBSTANCE ADDED BY NIELSON TO MODIFY THE EARLY CK DETERMINATION METHOD
ADDITION OF SULFHYDRYL COMPOUNDS AND AMP
EFFECT OF SULFHYDRYL COMPOUNDS AND AMP ON CK ACTIVITY IN ASSAY
ASSURE MAXIMUM CK ACTIVITY
DELAY ADENYLATE KINASE ACTIVITY
WHO PUBLISHED THE OPTIMIZED CONDITIONS FOR MEASURING CK
SZASZ IN 1976
SCANDINAVIAN COMMITTEE ON ENZYME
DIRECTION OF REACTION USED IN STANDARDIZED METHODS FOR CK DETERMINATION
REVERSE REACTION
ACTIVATION BY NAC
IFCC EXTENDED THE STANDARDIZED METHOD’S TEMPERATURE TO WHAT
37C
FORWARD REACTION OF CK
CREATINE PHOSPHATE + ADP —CK—> CREATINE + ATP
REVERSE REACTION OF CK
CREATINE + ATP —CK—> CREATINE PHOSPHATE + ADP
PH OF THE FORWARD REACTION OF CK
PH 9.0
PH OF THE REVERSE REACTION OF CK
PH 6.7
ACTIVATOR OF CK
MAGNESIUM
DIRECTION OF REACTION IN THE TANZER-GILBARG METHOD
DIRECT OR FORWARD
RELATIONSHIP BETWEEN THE RATE OF OXIDATION OF NADH AND CK ACTIVITY
DIRECTLY PROPORTIONAL
SPEED DIFFERENCES OF THE CK REACTION IN THE TANZER-GILBARG METHOD
REVERSE IS 5X FASTER THAN THE FORWARD REACTION
METHODS FOR THE SEPARATION AND QUANTITATION OF CK ISOENZYMES
ELECTROPHORESIS
IMMUNOASSAYS
COLUMN CHROMATOGRAPHY
ELECTROPHORETIC MIGRATION OF CK ISOENZYMES
CATHODE — MI : MM - MACRO - MB - BB — ANODE
IMMUNOASSAYS USED TO MEASURE CK-MB PROTEIN MASS
RADIO-IMMUNOASSAY
MONOCLONAL AB SPECIFIC FOR CK-MB
ASSOCIATIONS THAT CAN BE MADE WITH THE MEASUREMENT OF PROTEIN MASS
TYPE OF MYOCARDIAL INFARCTION
DEGREE OF DAMAGE
[DEGREE OF DAMAGE TO LEFT VENTRICLE]
[PROTEIN MASS VALUE]
SMALL MI
<10% DAMAGE
10-60 UG/L
[DEGREE OF DAMAGE TO LEFT VENTRICLE]
[PROTEIN MASS VALUE]
MEDIUM MI
10-30% DAMAGE
60-225 UG/L
[DEGREE OF DAMAGE TO LEFT VENTRICLE]
[PROTEIN MASS VALUE]
LARGE MI
> 30% DAMAGE
225 UG/L
REFERENCE VALUE OF CK IN IMMUNOASSAYS
UP TO 5 UG/L
[PRINCIPLE]
COLUMN CHROMATOGRAPHY
SEPARATION OF SUBSTANCES IN A COLUMN PACKED WITH AN ADSORBENT
FACTOR THAT AFFECTS THE MIGRATION OF SUBSTANCES IN COLUMN CHROMATOGRAPHY
AFFINITY OF EACH SUBSTANCE FOR THE ADSORBENT
SOLUTION INCORPORATED INTO THE COLUMN IN COLUMN CHROMATOGRAPHY
DIETHYL AMINO ETHYL EXCHANGE RESIN
PH USED IN COLUMN CHROMATOGRAPHY
PH 8.0
WHAT CHARGE BINDS THE GREATEST TO THE COLUMN
ISOENZYMES OF CK WITH A NET NEGATIVE CHARGE
SOLUTION USED TO ELITE OR REMOVE THE ISOENZYMES IN THE COLUM CHROMATOGRAPHY METHOD
SODIUM CHLORIDE
ISOENZYMES SEPARATED IN THE COLUMN CHROMATOGRAPHY CAN BE ASSAYED WITH WHAT METHOD
ROSALKI METHOD
DIRECTION OF REACTION IN THE FLUORESCENCE AND BIOLUMINESCENCE METHOD
REVERSE
CREATINE FORMED IN THE F&B METHOD IS ESTIMATED FLUOROMETRICALLY AFTER THE ADDITION OF WHAT SOLUTION
NINHYDRIN
SOLUTION USED TO FOLLOW THE RATE OF ATO FORMATION AND UTILIZATION IN THE F&B METHOD
LUCIFERIN
LUCIFERASE
IONS THAT INHIBIT CK ACTIVITY
EXCESS Mg, Cl, SULFATES
EFFECT OF EXCESS MG, CL, CA, AND SULFATES ON CK ACTIVITY
FLASE DECREASE
These inhibit CK activity
OTHER SUBSTANCES THAT INHIBIT CK ACTIVITY
IODOACETATE
ADP
CITRATE
FLUORIDE
L-THYROXINE
STABILITY OF CK ACTIVITY IN SERUM SAMPLES
2 HOURS, RT
8-12 HOURS, 4C
2-3 DAYS, FROZEN
EFFECT OF HEMOLYZED SAMPLES ON CK ACTIVITY
FALSE INCREASE
Side reactions due to AK, ATP, and G6PD
MAIN SOURCE OF AK
ERYTHROCYTES
PLATELETS
SKELETAL MUSCLE
BRAIN
LIVER
HEART
KIDNEY
SMALL INTESTINES
HOW DOES AK INTERFERE WITH CK ACTIVITY
AK CATALYZES THE DIRECT CONVERSION OF ADP TO ATP AND AMP
MORE ATP GENERATED INCREASES CK ACTIVITY
IN WHAT SAMPLES ARE LOWER CK VALUES OBSERVED
SAMPLES WITH HIGH ALP ACTIVITY
EFFECT OF LIGHT ON CK ACTIVITY
FALSE DECREASE
Light inactivates CK
EFFECT OF LIPEMIC SAMPLES ON CK ACTIVITY
FALSE INCREASE
High Abs readings
EFFECT OF INTRAMUSCULAR INJECTIONS ON CK ACTIVITY
FALSE INCREASE
Sedatives and analgesics are examples
[CK ACTIVITY DIFFERENCE]
MEN VS. WOMEN
MEN HAVE HIGHER VALUES
Due to muscle mass
[CK ACTIVITY DIFFERENCE]
NEWBORNS VS. ADULTS
NEWBORNS HAVE HIGHER VALUES
Due to skeletal muscle trauma during birth
EFFECT OF RECENT EXERCISE ON SERUM CK
FALSE INCREASE
Due to muscle activity
CK-MB may be increased when skeletal activity is sustained
FUNCTION OF MAGNESIUM
COFACTOR FOR THE MAXIMUM CATALYTIC ACTIVOTY OF THE ENZYME
FUNCTION OF ADENOSINE 5 MONOPHOSPHATE
INHIBITS ADENYLATE KINASE ACTIVITY
FUNCTION OF N-ACETYLCYSTEINE
PREVENTS OXIDATION OF SULFHYDRYL GROUP OF CK
WHAT MUST CK HAVE TO BE ACTIVE
A REDUCED SULFHYDRYL GROUP
When it is oxidized, no enzyme activity may be observed
FUNCTION OF EDTA
CHELATES CALCIUM
Ca inhibits CK activity
EFFECT OF TEMPERATURE ON CK ACTIVITY
CK ACTIVITY INCREASES
TEMPERATURES AT WHICH CK ACTIVITY ALTERS
Up to 45C — CK activity increases
> 45C — CK activity rapidly decreases
55C and above — CK activity is no longer detectable
REFERENCE METHOD FOR CK-MB ASSAYS
IMMUNO-INHIBITION METHOD BY WURZBURG
WHAT CK SUBUNIT INCREASES WHILE TOTAL CK REMAINS NORMAL
IN WHAT CLINICAL CONDITION DOES THIS OCCUR
CK-MB
MYOCARDIAL INFARCTION
[PRINCIPLE]
IMMUNO-INHIBITION METHOD BY WURZBURG
MEASURMENT OF CK ACTIVITY IN THE PRESENCE OF AN AB TO CK-M MONOMER
WHAT DOES THE AB TO CK-M MONOMER INHIBIT
INHIBITS CK-MB AND 1/2 OF CK-BB
METHOD USED TO QUANITATIVELY DETERMINE CK-B ACTIVITY
CK METHOD BY ROSALKI
RELATIONSHIP OF THE RATE OF NADH FORMATION AND CK-B ACTIVITY
DIRECTLY PROPORTIONAL
HOW TO COMPUTE FOR CK-MB
MULTIPLY CK-B ACTIVITY BY 2
[PRINCIPLE]
IMMUNOPRECIPITATION
ANTISERA AGAINST PURE CK-MM AND CK-BB IS USED TO QUANTITATE THE THREE CK ISOENZYMES
RESULT OF CK-MM AND BB ISOENZYMES IN IMMUNOPRECIPITATION
COMPLETELY PRECIPITATED
SUBSTANCE USED TO PRECIPITATE CK-MM AND BB
HOMOLOGOUS RABIT ANTISERA
RESULT OF CK-MB ISOENZYMES IN IMMUNOPRECIPITATION
PARTIAL PRECIPITATION
WHAT CK ISOENZYMES ARE NORMALLY PRESENT IN SERA OF HEALTHY PATIENTS
CK-MM
CK-MB
EFFECT OF CK-BB ACTIVITY ON CK-MB VALUES
INCREASED
Overestimation of CK-MB values
EFFECT OF MACRO-BB ACTIVITY ON CK-MB VALUES
INCREASE
Macro BB will be measured as B-subunit
VALUE OF CK-B IF MACRO BB IS SUSPECTED
MORE THAN 20% OF TOTAL CK ACTIVITY
TYPE OF CK-M ANTIBODY USED IN THE ASSAY
ANTI-HUMAN CK-M AB
GOAT
AMOUNT OF CK-MM INHIBITED BY GOAT SERUM
UP TO 1500 IU/L OF CK-MM
37C
AMOUNT OF CK-MM INHIBITED BY ANTI CK-MM AB OF JOCKERS WRETOU
100 % CK-MM
80% CK-MB
SIMILAR PROPERTIES OF INHIBITION BY THE ANTI CK-MM AB OF JOCKERS-WRETOU WERE FOUND IN WHAT OTHER ANTISERUM
RABBIT SERUM BY MORIN
[DISADVANTAGE]
USE OF MORIN’S RABBIT ANTISERA
RESIDUAL ACTIVITY IN THE SAMPLE AFTER ADDITION OF THE ANTISERUM IS TOO LOW
[ADVANTAGE]
USENOF ANTI HUMAN CK-M AB
GOAT ANTISERA
INHIBITS CK-MM TO 100% AND CK-MB TO 50+/55%