Creatine kinase isoenzyme MB assay by electrophoresis

A method for determination of the creatine kinase isoenzyme MB (CK-MB) is reported: separation of the isoenzymes was done by electrophoresis and the activity of the isoenzyme bands quantitated by scanning fluorometry. Total CK activity was used for calculation of CK-MB level. The precision of the method was satisfactory: coefficient of variation 5-10%. Its accuracy good: CK-MB was consistently found in high concentrations in tissue extracts of myocardium, but was virtually absent in skeletal muscle and could not be demonstrated in serum from patients with skeletal muscle damage. The sensitivity of the method fitted its clinical use: CK-MB was undetectable (less than 5 U/l) in normal sera, below 30 U/l in seventy-six out of seventy-seven patients in whom the diagnosis of acute myocardial infarction (AMI) was disproved, and above 30 U/l in all seventy-two patients with AMI according to WHO criteria. The CK-MB concentration in serum rises to a maximum about 20 h after onset of clinical symptoms of AMI and reaches baseline levels 20-30 h later. The electrophoretic CK-MB method is easy, fast and reliable and is considered as an important diagnostic test for AMI.     

Activity of serum aspartate aminotransferase isoenzymes in patients with acute myocardial infarction

Activities of aspartate aminotransferase (AST) isoenzymes were determined in serial serum samples from 40 cases of acute myocardial infarction, and compared with activities of creatine kinase, CK-MB isoenzyme, lactate dehydrogenase, and alpha-hydroxybutyrate dehydrogenase for temporal changes. Cytosolic (soluble) AST (s-AST) and mitochondrial AST (m-AST) respectively increased 6.6 and 9.0 h after onset of chest pain. The median time at which serum m-AST activity peaked (15.8 U/L, range 6.4-53.5 U/L) was 47.8 h after the onset of infarction, 19.8 h later than the peak s-AST activity (171 U/L, range 53-517 U/L) and m-AST also disappeared from the serum more slowly than s-AST (p less than 0.001). Serum m-AST values were above normal for at least six days after the infarct. The ratio of m-AST to total AST in serum increased after myocardial infarction, being greatest (20%, range 11-32%) on the third day after onset. For individuals, peak activities of s-AST correlated well with total CK (r = 0.91) and CK-MB (r = 0.86) peak activities, indicating that s-AST also reflects the infarct size. However, m-AST correlated poorly with the enzymes commonly used in infarct diagnosis; it apparently provides different biological information.     

Serum creatine kinase isoenzyme MB concentration after endomyocardial biopsy

Serum total creatine kinase (CK), CK-MB and myoglobin (Mb) were serially determined in 17 patients who underwent endomyocardial biopsy. Mean total CK levels increased from 36 +/- 27 U/l 30 min before biopsy to a maximum of 112 +/- 77 U/l 8 h following the procedure (p less than 0.05). Similarly, Mb concentrations rose from 57 +/- 55 micrograms/l to 119 +/- 57 micrograms/l 30 min after biopsy (p less than 0.05). Normalization of total CK and Mb levels occurred within 16 and 8 h, respectively. A new immunoenzymetric assay (IEMA) was used to measure the mass concentration of the CK-MB molecule. The initial CK-MB levels were 0.2 +/- 0.4 microgram/l; a small but significant elevation was recorded as early as 2 h after biopsy (1.6 +/- 1.5 micrograms/l, p less than 0.05). CK-MB returned to initial concentration 16 h after the beginning of the procedure. Comparison with the maximum CK-MB levels recorded in 16 myocardial infarction patients (258 +/- 172 micrograms/l, range 90-680 micrograms/l) indicated that the modest increase of CK-MB level detected after biopsy probably reflects a limited endomyocardium lesion at the sampling site, excluding any significant myocardial damage. Total CK and Mb, which showed more pronounced elevations than CK-MB, are likely to originate from other sources than the myocardium.     

Human myocardial and skeletal muscle enzyme activities: creatine kinase and its isozyme MB as related to citrate synthase and muscle fibre types

Summary. Activities of myocardial and skeletal muscle total creatine kinase (CK) and its isozyme MB were related to the oxidative capacity [measured as the citrate synthase (CS) activity] and to the contractile characteristics (estimated as the percentage of type I muscle fibres). Skeletal muscle biopsies were obtained both from physically trained and untrained men and myocardial biopsies from patients subjected to open-heart surgery performed because of mitral or aortic valve disease. Enzyme activities were determined on freeze-dried muscle specimens. The CK-MB activity was about twice as high in trained skeletal muscle as in untrained ones reaching the myocardial level. The total CK activity was about three times higher in skeletal muscle than in myocardium; the myocardium, however, had CS activity 3–4 times larger than that of skeletal muscle. A close correlation was demonstrated between activities of CK-MB on one hand and CS (r= 0·76) or percentage type I fibres (r= 0·83) on the other hand suggesting a connection between CK-MB activity and the oxidative capacity of the cell. This was in contrast to total CK where different regressions were obtained when comparing the myocardium and the skeletal muscle of trained or untrained men. In conclusion, CK-MB activity in trained skeletal muscle in athletes were similar to that in myocardium. CK-MB was related to the oxidative capacity and formation of cellular energy in skeletal and heart muscle.     

Serum creatine kinase and CK-MB isoenzyme responses to acute and prolonged swimming in trained athletes

Six highly-trained male swimmers completed a maximum work capacity tethered swim and a 1-h continuous tethered swim at approximately 70% VO2max in order to evaluate total serum creatine kinase and CK-MB isoenzyme changes. Venous blood obtained before, 5 min post-, 6 h post-, and 24 h post-exercise was analyzed for total serum CK (kinetic UV method, normal = less than 100 U/l) and CK-MB isoenzyme (quantitative electrophoretic technique, normal = less than 5 U/l). VO2max averaged 4.59 +/- 0.28 l/min, with a mean total work time of 24.5 min to achieve maximum capacity. Mean resting total CK was 100.5 +/- 15.8 U/l. Compared to rest, neither swim bout produced a significant (p greater than 0.05) elevation in mean total creatine kinase. No CK-MB isoenzyme was observed in any post-exercise blood sample. Swimming, performed by highly-trained swimmers at high levels of intensity or for prolonged durations, may not impose sufficient degrees of trauma producing muscular stress. Therefore, the structural integrity of the cell membrane is maintained and the loss of intracellular creatine kinase to the bloodstream prevented.     

Creatine kinase:aspartate aminotransferase activity ratio as an indicator of the source of an increased creatine kinase activity

Although measurements of creatine kinase isoenzyme 2 (CK-MB) are often used to diagnose acute myocardial infarction, their sensitivity and specificity are less than 100%. Because skeletal muscle contains more CK and less aspartate aminotransferase (AST) than cardiac muscle, the CK/AST ratio might provide a useful adjunct in evaluating the source of a supranormal value for CK. I established the following decision levels in a retrospective study of 342 patients: ratios less than 14 (if total CK was 300-1200 U/L), less than 20 (CK 1201-2000 U/L), or less than 25 (CK greater than 2000 U/L) suggested myocardial infarction, with a sensitivity of 95% and a specificity of 65%. In a validation study with 277 additional patients, liver disease and alcohol abuse caused erroneous results, leading to exclusion of 22% of these patients. In the remaining cases, sensitivity was 94%, specificity 90%. The CK/AST ratios changed little with time, suggesting that a single value would be adequate for evaluating patients with increased CK.     

Immunoenzymetric assay for creatine kinase MB with subunit-specific monoclonal antibodies compared with an immunochemical method and electrophoresis

Results of an immunoenzymetric assay (TANDEM-E CKMB) for creatine kinase (CK; EC 2.7.3.2) MB isoenzyme, in which subunit-specific monoclonal antibodies are used, were compared with those by an immunochemical method (Isomune-CK) and electrophoresis (Corning agarose gel). The study involved 200 patients; greater than 500 samples were analyzed by all three methods. The analytical performances were acceptable. Between-method correlation coefficients ranged from 0.881 to 0.975. Two reference intervals were established for the immunoassays: 0-4 micrograms/L (TANDEM) and 0-4 U/L (Isomune) for "normal" patients; 0-9 micrograms/L (TANDEM) and 0-14 U/L (Isomune) for noninfarct patients. Agreement with respect to increased CK-MB as defined by the reference intervals for the noninfarct patient was 96% between TANDEM and electrophoresis, 90% between Isomune and electrophoresis. All three methods are acceptable for use in determining CK-MB, but the overall diagnostic efficiencies for the mass or activity concentration of the isoenzyme and for its proportion of total CK activity, based on the predictive value model, are 92% (electrophoresis, 0-7 U/L), 90% (electrophoresis, 0-4%), 92% (TANDEM, 0-9 micrograms/L), 88% (TANDEM, 0-3% index), 88% (Isomune, 0-14 U/L), and 83% (Isomune, 0-4%). All three methods can detect CK-MB in serum, but its presence is not necessarily diagnostic of acute infarct. We recommend using the actual concentration of CK-MB to evaluate patients with suspected acute myocardial infarct, and the percentage of CK-MB when total CK is very high.     

Diagnostic use of CK-MM and CK-MB isoforms for detecting myocardial infarction

Following acute myocardial infarction, total CK and CK-MB levels begin to rise 5 to 6 hours after the onset of chest pain. The serial profile of the rise and fall of both activities is nearly always indicative of AMI. The recent increase in the use of thrombolytic agents in an attempt to attain reperfusion of occluded coronary arteries alters the enzyme profiles observed in blood after AMI. After successful reperfusion a washout phenomenon occurs in which early restoration of blood flow to damaged myocardium causes an early rise in total CK and MB levels above the normal range 2 to 4 hours after AMI, with earlier and higher peak enzyme values. Recently reports have appeared describing numerous serum and plasma CK-MM and CK-MB isoform patterns after AMI. Following release from injured myocardium CK-MM3 and CK-MB2 (designated the tissue isoforms) are converted in the circulation to post-translation products (MM2, MM1, MB1, respectively). Studies have now shown that CK-MM isoform patterns provide a unique means of assessing the time of onset of necrosis and a monitor of the duration of enzyme release from the site of injury. Following AMI, MM3, the MM3/MM1 ratio, or both rises and peaks earlier than either total CK or CK-MB levels. During successful reperfusion, the rate of rise of CK-MM3 is more rapid and the MM3/MM1 ratio peaks earlier than without reperfusion. However, any concomitant release of CK-MM3 from skeletal muscle would decrease the clinical utility of MM isoforms in detecting myocardial damage. Recent advances in technology have shown that CK-MB2 rise parallels the CK-MM increase and also rises earlier than total CK and total MB levels and provides increased specificity for the myocardium. The full potential of the diagnostic utility of MM and MB isoforms will not be realized until a reliable, sensitive, simple, and rapid quantitative assay becomes available.     

Mitochondrial and cytoplasmic isoenzymes of aspartate aminotransferase in sera of patients after myocardial infarction

Mitochondrial and cytoplasmic isoenzymes of aspartate aminotransferase (AST) were studied in the sera of 42 patients following acute myocardial infarction and compared to creatine kinase (CK), lactate dehydrogenase (LDH) and alanine aminotransferase (ALT). Mitochondrial AST( ASTm ) was detected in 93% (39/42) of patients. Maximum recorded ASTm activity was 59.5 +/- 8.8 U/l and was found 39.4 +/- 3.5 hours after the onset of symptoms (chest pain) of myocardial infarction. In contrast the maximum recorded cytoplasmic AST ( ASTc ) activity was greater (327 +/- 23 U/l) and it occurred earlier (33.5 +/- 2.2 hours) after onset of infarction compared to ASTm . ASTm correlated significantly (p less than 0.05) with ASTc , LDH and ALT but not with total CK or CK-MB. ASTc correlated significantly (p less than 0.05) with total CK, CK-MB and LDH but not ALT. Maximum recorded ASTm activity was significantly associated with the clinical assessment of left ventricular failure ( Killip classification) but not with ventricular arrhythmias. In a subset of 15 patients evaluated with invasive hemodynamic measurements of cardiac output and pulmonary capillary wedge pressure. ASTm correlated significantly (p less than 0.05) and better than CK-MB with the hemodynamic assessment of left ventricular dysfunction. Thus ASTSm can be readily identified in sera of patients after acute myocardial infarction and may be of value in the evaluation of patients with acute myocardial infarction.     

Clinical effectiveness of the Du Pont aca measurement of creatine kinase MB in serum from patients in a coronary-care unit

We evaluated the clinical effectiveness of measuring creatine kinase (CK; EC 2.7.3.2) isoenzyme MB and lactate dehydrogenase (LD; EC 1.1.1.27) isoenzymes in diagnosis of acute myocardial infarction. We used an agarose electrophoresis method to measure CK and LD isoenzymes and the Du Pont aca column method to measure CK-MB. Serial blood specimens were drawn from 100 patients consecutively admitted to our Coronary Care Unit. Because of the low diagnostic specificity for CK-MB measurements by both agarose electrophoresis and the discrete-analysis method, as compared with reported values, we re-evaluated our isoenzyme data by using Receiver Operating Characteristic curves. Such analysis of the data established optimal decision levels of greater than or equal to 25 U/L and greater than or equal to 18 U/L plus greater than or equal to 6% of total CK for serum CK-MB measured by the agarose electrophoresis and the aca methods, respectively, and an optimal decision level of greater than or equal to 0.92 for the ratio of LD 1/2 measured after agarose electrophoresis. At these decision levels we obtained a sensitivity of 100%, 100%, and 95% and a specificity of 94%, 92%, and 90% for CK-MB (agarose electrophoresis), CK-MB (aca), and the LD 1/2 ratio, respectively.     

Diagnosis of perioperative myocardial infarction by considering relationship of postoperative electrocardiogram changes and enzyme increases after coronary bypass operation

We report the results of enzyme determinations in sera from 88 patients, 65 of whom showed inconspicuous reconvalescence, 14 who had myocardial infarction within 24 h (MI 1) after bypass surgery, and nine with myocardial infarction between 24 and 48 h postoperatively (MI 2). We wanted to determine whether the consequent measurement of activities of total creatine kinase (CK), CK isoenzyme MB (CK-MB), lactate dehydrogenase, alpha-hydroxybutyrate dehydrogenase, and aspartate aminotransferase, conducted as a part of routine laboratory diagnostics, provided meaningful information for diagnosing infarcts besides that obtained from the electrocardiogram. The postoperative mean values of the enzyme activities in blood were significantly different among the three groups; however, only a combined evaluation of CK and CK-MB by means of a discriminant analysis allowed the prediction of MI (sensitivity: MI 1 = 98.5%, MI 2 = 95.4%; specificity: MI 1 = 71.4%, MI 2 = 81.8%). CK greater than 600 U/L or CK-MB greater than 45 U/L supports the diagnosis of acute MI.     

Comparison of ASAT, CK, CK-MB, and LD for the estimation of acute myocardial infarct size in man

The purpose of this study was to set up a simple and reliable procedure for estimating acute myocardial infarct (AMI) size by measuring serum enzymes in a few daily blood samples. Peak enzyme values and estimated infarct size from one, two, or three daily samples of aspartate aminotransferase (ASAT), creatine kinase (CK), CK-MB, and lactate dehydrogenase (LD) were compared with the extent of myocardial necrosis measured at autopsy in 22 patients who died from AMI. The correlation between the extent of the necrosis measured and peak serum enzymes from one daily blood sample was highest for CK-MB (r = 0.78) and LD (r = 0.73) compared to CK (r = 0.68) and ASAT (r = 0.67). To obtain a significant correlation, however, two patients had to be excluded from the ASAT and LD analyses. No significant improvement was obtained by more frequent blood sampling. Estimation of infarct size did not improve the correlation significantly for any enzyme, although the coefficient of correlation for CK-MB increased slightly (r = 0.83). Serum CK-MB determination provides a semiquantitative estimate of infarct size, but the other enzymes may give erroneous estimates owing to lesser cardiospecificity.     

肌酸激酶同工酶质量判定肌病心肌损害的局限性

目的 通过对比小儿心肌炎、肌病时血清中肌酸激酶同工酶(CK-MB)质量与肌钙蛋白I(cTnI)和肌红蛋白(Mb)的动态变化,观察CK-MB质量在判定心肌损伤中的意义.方法 测定40例心肌炎患儿(其中有20例为暴发性心肌炎)和38例肌病患儿的肌酸激酶(CK)、CK-MB活性、CK-MB质量、cTnI、Mb、心电图以及脉冲多普勒超声心动图;肌病组同时进行肌电图、遗传代谢病筛查以及基因检测.以同期本院发育儿科门诊除外甲状腺功能减低症的10例身矮待查儿童为对照.结果 ①健康对照组儿童CK(U/L)为95.0±27.0,CK-MB活性(U/L)为22.6±1.3,CK-MB质量(μg/L)为2.4±0.3,cTnI(μg/L)为0.012±0.001.②心肌炎组患儿治疗前CK(1 033.0±408.0)、CK-MB活性(101.2±31.5)、CK-MB质量(38.2±13.2)、cTnI(5.544±1.554)均较健康对照组明显升高(均P＜0.01);随着治疗时间延长,各项指标逐渐下降;治疗2周后CK(59.3±25.1)、CK-MB活性(24.6±13.2)、CK-MB质量(3.3±2.9)、cTnI(0.125±0.128)均恢复至正常水平(均P＞0.05).治疗1周后CK、CK-MB质量增高率即较治疗前明显下降[CK:5.9％(1/17)比56.4％(22/39);CK-MB质量:8.3％(1/12)比61.1％(22/36),均P＜0.01],CK-MB质量恢复先于cTnI,增高率出现明显差异[8.3％(1/12)比73.7％(14/19),P＜0.05].③肌病组治疗前CK(10 193.0±1 447.0)、CK-MB活性(311.7±44.4)以及CK-MB质量(229.2±47.9)均较健康对照组明显升高(均P＜0.01),但cTnI不高(0.021±0.002);治疗2周后CK(5 735.6±6 187.8)、CK-MB活性(170.7±143.0)、CK-MB质量(207.4±136.6)仍维持在高水平,cTnI(0.230±0.150)则维持在正常水平;各项指标的增高率与治疗前均无显著差异[CK:85.7％(6/7)比97.4％(37/38); CK-MB活性:85.7％(6/7)比97.4％(37/38);CK-MB质量:100.0％(2/2)比94.1％(32/34);cTnI:0(0/1)比6.4％(2/31),均P＞0.05].结论 ①在心肌炎时,CK-MB质量与cTnI一致,急性期升高,恢复期降至正常,但CK-MB质量观察窗短于cTnI.②在肌病时,CK-MB质量与cTnI 分离,前者治疗前后均升高,后者正常,故用测定CK-MB质量来判定肌病患儿是否有心肌损害意义有限.     

Prediction of infarct size from serial CK determinations: evaluation by clinical studies and computer simulation.

To assess reduction of infarct size by therapeutic intervention, a high predictive accuracy is mandatory. The CK release in the circulation (CKr) was studied in 12 consecutive patients after uncomplicated myocardial infarction, admitted within 5 h after onset of symptoms. Despite improvement of existing methods, such as a more frequent sampling, CK-MB determination instead of total CK determination and use of a gamma-exponential instead of a log-normal curve-fitting technique, the correlation between CKr predicted from measurements within 7 h after the start of CK rise and CKr calculated after completion of the CK curve remained poor. Computer simulations were done to investigate measurement errors as a cause of this failure. Normally distributed noise, with standard deviations ranging from 0.2% to 8.0% of peak CK-MB, was added to the first points of an ideal gamma-exponential CK-MB curve and predictions were made from these "noisy" points. A small noise already produced a great variation in prediction: 0.8% noise resulted in a deviation of predicted CKr from calculated CKr ranging from --20 to +6%. It is concluded that adequate prediction of infarct size from serial CK determinations in the first 7 h after onset of the CK rise must fail if the precision of the biochemical determination is not less than 0.4%.     

Creatine kinase isoenzymes

Creatine kinase (CK), a widely distributed enzyme in the body, has its highest activities in skeletal muscle and myocardium; when serum CK activities are abnormally increased, injury to these organs must be part of the differential diagnosis. The isoenzyme CK-MB is the most important biochemical test in the diagnosis of acute myocardial infarction.     

Creatine kinase MB isoforms in patients with skeletal muscle injury: ramifications for early detection of acute myocardial infarction.

We measured total creatine kinase (CK), CK-MB isoenzyme, and the MB isoforms in 202 serum and plasma samples from nine groups of patients and normal individuals: 39 with acute myocardial infarction (MI), divided according to time between the onset of chest pain and blood collection (1-6 h, 7-12 h, and 13-48 h); 26 with chest pain for whom an MI was ruled out, sampled at admission; 17 undergoing bypass surgery or cardiac catheterization, sampled within 6 h after either procedure; 17 with acute skeletal muscle injury, sampled within 8 h after injury; 30 marathon runners immediately after a race; 17 runners and other athletes > 12 h after training or a race; 12 with cerebral injury or seizures, sampled at admission; 8 with closed head injury, sampled at admission; and 38 normal subjects. CK-MB (relative index) and MB isoforms (MB2/MB1) were respectively increased in 15% and 75% of MI patients 1-6 h after onset, 94% and 94% after 7-12 h, and 88% and 8% after 12 h, and in 87% and 82% of cardiac surgery patients. MB isoforms were increased in most patients with acute skeletal muscle trauma and in subjects examined after exercise, but were within normal limits in patients for whom MI was ruled out, patients with cerebral trauma, and normal individuals. The relative index of MB/total CK was normal in essentially all individuals in the last groups, including those with acute skeletal muscle trauma. We concluded that the CK-MB isoform ratio is increased in both acute skeletal muscle injury and MI. The isoform ratio is most useful for distinguishing recent from old (> 12 h) injury.     

Cardiac troponin I and troponin T: recent players in the field of myocardial markers.

The troponin (Tn) complex consists of three subunits referred to as TnT, TnI and TnC. Myocardium contains TnT and TnI isoforms which are not present in skeletal muscles and which can be separated from the muscular isoforms by immunological techniques. Using commercially available immunoassays, clinical laboratories are able to determine cardiac TnT and TnI (cTnT and cTnI) quickly and reliably as classical cardiac markers. After acute myocardial infarction, cTnT and cTnI concentrations start to increase in serum in a rather similar way than CK-MB, but return to normal after longer periods of time (approximately one week). Because of their excellent cardiac specificity, Tn subunits appear ideally suited for the differential diagnosis of myocardial and muscular damage, for example in noncardiac surgery patients, in patients with muscular trauma or with chronic muscular diseases, or after intense physical exercise. cTnT and cTnI may also be used for detecting evidence of minor myocardial damage: therefore they have found new clinical applications, in particular risk stratification in patients with unstable angina. In spite of the possible reexpression of cTnT in human skeletal muscles, and of the lack of standardization of cTnI assays, Tn subunits are not far to meet the criteria of ideal markers for acute myocardial injury. Only an insufficient sensitivity in the first hours following the acute coronary syndroms requiries to maintain an early myocardial marker in the cardiac panel for routine laboratory testing.     

Extent of myocardial tissue damage during transmyocardial laser revascularization with the CO2 Heart Laser.

OBJECTIVE AND BACKGROUND: Transmyocardial laser revascularization (TMR) is the only surgical treatment for patients with severe diffuse coronary artery disease, who are not candidates for bypass grafting or percutaneous angioplasty. However, vaporization of tissue during the creation of channels leads to a certain loss of viable myocardium during every TMR procedure. METHODS: We analyzed serum levels of creatine kinase and creatine kinase MB subtype in 163 patients after sole TMR with a CO2 laser (wave length 10.6 microm, 800-watt power). The control group consisted of 35 consecutive CABG patients and 30 consecutive redo-CABG patients. Additionally, in the TMR group we measured echocardiographically the left ventricular ejection fraction before and after TMR. We recorded the total amount of laser energy applied, average and maximum energy per channel, and the number of created channels, in order to calculate the correlation between these parameters and postoperative enzyme levels or changes in the LVEF. RESULTS: After TMR, we measured higher creatine kinase levels compared to those in CABG patients (607.8+/-558.4 U/L vs. 285.0+/-292.3 U/L, p < 0.01). The relative proportion of CK-MB of total CK, however, was significantly lower after TMR, compared to that of the control group (4.5+/-3.0% vs. 10.1+/-6.4%, p < 0.01). Patients with a pronounced postoperative increase in CK-MB levels or a higher percentage of CK-MB of total CK also after TMR operations show a decline in left ventricular contractility. In the laser group, the maximum enzyme levels were detected significantly later than in the control group (25.0+/-19.4 h postoperatively vs. 8.7+/-9.1 h, p < 0.01). There was no significant correlation between the technical laser parameters or the number of created channels and the percentage of CK-MB of total CK or changes in left ventricular ejection fraction. CONCLUSIONS: CO2 laser TMR does not result in significant injury to the myocardium. Cardiac enzymes play an important role in the detection of perioperative myocardial infarction in TMR patients.     

Release patterns of CK-MB and mitochondrial CK following myocardial ischaemia

Following myocardial damage as in acute myocardial infarction (AMI) or open heart surgery, the tissue damage might result in a release of mitochondrial CK (CK-MIT). The presence of this CK isoenzyme in serum may be detected after chromatographic separation of CK-activity on Sephacryl S-200. By combining chromatographic separation of CK-MB with immunologic inhibition of CK-M, both CK-MB and CK-MIT can be estimated in serum. Using this procedure changes in enzyme activities were studied in ten patients with AMI and twelve patients subjected to open heart surgery using cardioplegia. Following AMI CK-MB peaked about 24 h after onset of ischaemic symptoms. CK-MIT increased similarly and reached a plateau after 24 h where it remained during an additional 24-36 h. At peak CK-MB concentration, the corresponding CK-MIT activity was about 22% of the CK-MB activity. Following cardiac surgery there was a rapid release of CK-MB with a peak about 5 h after release of aortic cross-clamping, and with a simultaneous CK-MIT activity amounting to 19% of the CK-MB activity. In conclusion, CK-MIT is released into serum following myocardial ischaemia. Its appearance has time characteristics similar to that of other mitochondrial enzymes. The CK-B method does not specifically determine CK-B, but non-CK-M, which in cardiac ischaemia at peak serum CK-MB concentrations includes about 20% CK-MIT.     

Simultaneous automated measurement of serum total CK and CK-MM lsoform ratio in serum

We automated a two-step kinetic procedure for determining serum CK-MM isoform ratio using an immunoinhibition method. By measuring the total CK activity and the residual CK activity (serum CK-MM isoform) remaining after the inhibition by tissue CK-MM isoform specific monoclonal antibody reagent (CK-M01) the CKMM isoform ratio is calculated using the difference between total CK and residual CK activities divided by the residual CK activity. Linearities of total CK and residual CK assays were?7750 U/L and 2,500 U/L, respectively; within-run CVs of isoform ratio (N = 10) were 2.8 and 7.0% (mean 0.14 and 0.60), respectively. The MM³/MM¹ isoform ratio obtained with the proposed method (X) correlated well with the results of electrophoretic method (Y) according to the equation: Y = 0.98X ?0.3, r = 0.988. The normal reference range of isoform ratios obtained by assaying 1,222 serum samples from healthy subjects was 0.09–0.75. The isoform ratio increased after onset of chest pain, peaking at 2–6 hr thereafter. A mean isoform ratio of 1.86 was obtained with serum sample from 86 patients diagnosed as having an acute myocardial infarction (AMI). This method is accurate and highly sensitive, as the detection and early diagnosis of AMI can be completed in 10 min. © 1994 Wiley-Liss, Inc.