Ameliushkina V A, Krauze E G, Rabich G, Cherniad'eva I F, Titov V N
Lab Delo. 1991(1):17-22.
A method for measuring blood serum glycogen phosphorylase (GP) activity is described, informative at early stages of myocardial infarction. The method is sensitive and available for clinical biochemistry laboratories. It consists in preliminary purification of GP from serum proteins and metabolites by affinity chromatography in micro-columns and subsequent measurement of the activity in the eluate. The procedure involves selective GP sorption on starch, washing, and subsequent desorption with glycogen solution. GP activity is measured by the kinetic spectrophotometric technique, based on enzymic measurement of glucose-1-phosphate, the product of glycogen consumption reaction, at a wavelength of 340 nm. Conditions of serum GP chromatographic purification are modified in the suggested procedure, this improving the sensitivity of the enzyme measurement. Blood serum GP activities were measured in patients with various cardiac diseases--myocardial infarction (15 cases), angina of rest and effort (53), essential hypertension (30). Different methods of GP activity measurements are considered. Recommendations on the use of the described method, a sensitive test for the diagnosis of myocardial infarction, are given.
本文描述了一种测定血清糖原磷酸化酶(GP)活性的方法,该方法在心肌梗死早期具有重要意义。该方法灵敏,临床生化实验室均可采用。它包括通过微柱亲和色谱法从血清蛋白和代谢物中初步纯化GP,随后测定洗脱液中的活性。该过程包括GP在淀粉上的选择性吸附、洗涤,以及随后用糖原溶液解吸。GP活性通过动力学分光光度技术测定,基于在340nm波长下对糖原消耗反应产物葡萄糖-1-磷酸的酶促测定。在建议的方法中,血清GP色谱纯化条件有所改进,从而提高了酶测定的灵敏度。对患有各种心脏病的患者——心肌梗死(15例)、静息型和劳力型心绞痛(53例)、原发性高血压(30例)——测定了血清GP活性。文中还考虑了不同的GP活性测量方法。给出了关于使用所述方法(一种诊断心肌梗死的灵敏检测方法)的建议。