糖原磷酸化酶同工酶BB测定方法的建立及其在急性冠脉综合征诊治中的价值

Assay of glycogen phosphorylase isoenzyme BB and its significance of diagnosis of ischemic myocardial injury

关键词:  糖原磷酸化酶  酶联免疫吸附法（ELISA）  急性冠脉综合征  早期诊断

Key words:  glycogen phosphorylase  ELISA  acute coronary syndrome(ACS)  early diagnosis

摘要:

［目的］分离纯化糖原磷酸化酶同工酶BB(GPBB)，建立血清GPBB定量测定方法，探讨其在急性冠脉综合征早期诊断及治疗中的价值。［方法］ 应用纯化GPBB抗原及其抗体建立GPBB定量测定方法。对34例健康人、28例不稳定心绞痛(UAP)患者和29例急性心肌梗死(AMI)患者分别进行血清GPBB、CK、CK-MB及cTnI测定。［结果］ GPBB被纯化100倍，比活性384 μmol/mg·min-1，回收率28%。建立了GPBB定量测定方法，确定抗原检测线性范围为0.3～20 ng。在AMI发作6 h内，患者血清中（n=26）GPBB浓度超过正常上限值7.4 μg/L。 GPBB诊断灵敏度是9.7%，明显高于CK、CK-MB（24.1%，20.8%，p<0.01）,与cTnI相当。AMI患者血清GPBB峰值水平(66.38±27.41μg/L)明显高于健康人（2.57±1.61) μg/L，p<0.01。28例UAP患者中21例血清GPBB升高≥7.4 μg/L，其均值是(23.2±14.2) μg/L，明显高于健康人和稳定心绞痛患者血清GPBB水平（p<0.01），与cTnI诊断阳性率相似，而UAP患者血清中CK、CK-MB均无明显升高。其中GPBB升高组（≥7.4 μg/L）4周内发生心性事件远高于GPBB<7.4 μg/L组（47.6%，14.3%，p<0.01）。AMI患者血清GPBB到达峰值时间早于CK和CKMB达到峰值时间。［结论］GPBB是AMI发作后6 h内敏感特异的生化指标，对判断UAP预后有一定的临床价值。

Abstract:

［Objective］ To establish a method of glycogen phosphorylase isozymes (GPBB) mass measurement and evaluate GPBB as a cardiac mark during the early diagnosis of acute myocardial infarction(AMI) and unstable angina pectoris(UAP). ［Methods］ GPBB and its antibody were purified. Establish a developed ELISA assay using polycolonal antibody. The mass of GPBB, the activity of CK, CKMB and cTnI in the serum obtained from 34 normal individuals, 14 stable angina pectoris patients, 28 UAP patients and 29 AMI patients were determined by ELISA, respectively. ［Results］ GPBB was purified 100 times with a resulting specific activity of 384 units per mg protein and with a 28% recovery. The calibration curve of the GPBB in serum is linear throughout 0.3 to 20 μg/L. GPBB in 26 AMI patients increased above 7.4 μg/L (URL) within the first 6h after AMI onset. The sensitivity of GPBB detected was 89.7%, which was higher than that of CK (24.1%, p<0.01) and CKMB (20.8%, p<0.01), equal to that of cTnI. The peak value of GPBB in AMI patients (66.38±27.41) μg/L was significantly higher (p<0.01) than that in normal individuals (2.57±1.61) μg/L. GPBB in 21 out of 28 UAP patients increased above 7.4 μg/L (URL) after chest pain onset. The peak value of GPBB in UAP patients (23.2±14.2) μg/L was significantly higher (P<0.01) than that in normal individuals (2.57±1.61) μg/L and stable angina (2.78±0.94) μg/L. No significant difference was found in serum CK and CK-MB among the 28 UAP patients. GPBB in 21 out of 28 UAP patients increased above 7.4 μg/L (URL) after chest pain onset. No significant difference was found in serum CK and CK-MB among UAP and stable angina groups. Among the 28 UAP patients, the incidences of cardiac events within four weeks were significantly higher in group II(GPBB>7.4 μg/L) than in group I (GPBB<7.4 μg/L ) (47.6% vs 14.3%，p<0.01).The time to the peak value of GPBB in patients with AMI was earlier than that of CK and CKMB. ［Conclusion］ GPBB is more sensitive than CK and CKMB in the patients within 6h after AMI onset. It is also a very useful cardiac mark in early diagnosis of AMI and UAP.