Everse J, Reich R M, Kaplan N O, Finn W D
Clin Chem. 1975 Aug;21(9):1277-81.
Lactate dehydrogenase isoenzymes can be distinguished kinetically by the fact that isoenzyme H is strongly inhibited a few seconds after the reaction is started if high concentrations of pyruvate are present, in contrast to the M isoenzyme. A new instrument that exploits this fact can measure both the total activity and the proportion of H isoenzyme in serum or plasma in 8 to 10 s. The instrument consists of a simplified stopped-flow apparatus in which the plasma is assayed for lactate dehydrogenase activity, and an electronic device that measures the rate of the reaction at two pre-set time intervals. The first rate is taken between 0.2 and 0.4 s after the reaction is started, a time at which both isoenzymes are fully active, and at which the rate obtained thus reflects total lactate dehydrogenase activity in the plasma sample. The second rate is measured 4 to 6 s after the start of the reaction, at which time the H isoenzyme has become inhibited and the observed rate compared to the initial rate is therefore proportional to the percentage of H isoenzyme activity in the serum. These two rates are electronically displayed on two three-digit voltmeters, the first display being the total activity, the second a number proportional to the inhibited slope. The percentage of M isoenzyme can then be calculated from the initial and final rate. A total of five to six repeat assays may be done within a minute on 1 ml of plasma or serum. This instrument may be of significant value in following the progress of myocardial infarctions and other diseases.
乳酸脱氢酶同工酶可以通过动力学方法加以区分,因为如果存在高浓度的丙酮酸,同工酶H在反应开始后几秒就会受到强烈抑制,这与M同工酶不同。一种利用这一原理的新型仪器能够在8到10秒内测定血清或血浆中乳酸脱氢酶的总活性以及H同工酶的比例。该仪器由一个简化的停流装置和一个电子设备组成,在停流装置中测定血浆中的乳酸脱氢酶活性,电子设备则在两个预设的时间间隔测量反应速率。第一个速率在反应开始后的0.2到0.4秒之间测定,此时两种同工酶均具有完全活性,所获得的速率反映了血浆样品中乳酸脱氢酶的总活性。第二个速率在反应开始4到6秒后测量,此时H同工酶已被抑制,因此观察到的速率与初始速率相比与血清中H同工酶活性的百分比成正比。这两个速率以电子方式显示在两个三位电压表上,第一个显示的是总活性,第二个显示的是与抑制斜率成正比的数字。然后可以根据初始速率和最终速率计算出M同工酶的百分比。在1毫升血浆或血清上,一分钟内总共可以进行五到六次重复测定。该仪器在跟踪心肌梗死和其他疾病的进展方面可能具有重要价值。