摘要
目的建立胶体金免疫层析检测人糖原磷酸化酶同工酶BB(GPBB)的方法。方法采用胶体金标记抗单克隆GPBB抗体D1,将其与生物素化的抗GPBB单抗D2包被于吸水纤维,将链霉亲合素结合于硝酸纤维素膜,制成免疫层析试条,依据试剂条上出现肉眼可见的红色线条判定结果。用其检测96例发病6h内的胸痛患者血浆中的GPBB,与心肌肌钙蛋白I(cTnI)和肌酸激酶MB同工酶(CK-MB)的检测结果比较,评价其诊断急性心肌梗死(AMI)的敏感性和特异性。结果金标记试条检测GPBB的浓度低限值为7.4ng/ml,与ELISA方法比较符合率达96.15%;诊断发病4h内和6h内AMI的敏感性分别为64.71%和83.92%,高于cTnI(29.42%、51.79%)和CK-MB(20.59%、55.36%),分别P<0.05,特异性比较差异无统计学意义。结论本法简便、快速、准确,可用于AMI早期筛查。
Objective To develop a simple, rapid, sensitive assay for the detection of human glycogen phosphorylase isoenzyme BB （GPBB） by colloidal gold immunochromatography. Methods The GPBB monoclonal antibody D1 was conjugated with colloid gold, and immobilized onto a dry membrane with blotinylated monoclonal antibody D2 together. Streptavidin was coupled with the nitrocellulose strip to prepare the test apparatus. The results were analyzed according to the red llne on nitrocellulose membrane. The rapid one-step immunochromatography was constructed and used for the detection of plasma GPBB from 96 patients with suspected AMI in 6 hours after onset of chest pain. The diagnostic validity was evaluated in terms of sensitivity and specificity compared with those of cardiac troponin-I （cTnI） and creatine klnase-MB （CK-MB）. Results The lowest limit of the immunochromatographic assay for detection of GPBB was 7. 4 ng/ml. Compared with the results obtained by ELISA, the coincidence rate of the two assays was 96. 15 %. The sensitivity of GPBB for AMI diagnosis was 64. 71% within 4 hours and 83.92% within 6 hours after onset of chest pain, which were significantly higher than those of cTnI （29.42% and 51.79%） and CK-MB （20. 59% and 55.36%） respectively. There was no significant difference in specificity between the two assays. Conclusion The simple, rapid and sensitive immunochromatographic assay for detection of GPBB may be applicable in the early diagnosis of AMI.
出处
《福建医药杂志》
CAS
2007年第5期119-120,共2页
Fujian Medical Journal
基金
福建省自然科学基金资助项目(C0210024)
福建省卫生厅青年科学研究基金项目(2001-1-16)
关键词
糖原磷酸化酶同工酶
急性心肌梗死
胶体金
免疫层析法
Human glycogen phosphorylase isoenzyme BB
Acute myocardial infarction
Colloidal gold
Immunochromatography
