【摘要】 肌酸激酶同工酶(Creatine kinase isoenzyme,CK-MB)是早期诊断急性心肌梗死(acute myocardial infarction,AMI)的重要标志物,该研究以荧光微球为标记物研究快速、灵敏的CK-MB荧光免疫层析检测方法。通过N-羟基琥珀酰(NHS)和1-乙基-3-(3-二甲氨基丙基)碳二亚胺盐酸盐(EDC)将荧光微球与市售的抗体共价偶联,优化抗体与荧光微球的偶联条件,对偶联获得的抗体荧光标记物进行表征,制备了CK-MB的荧光免疫层析检测卡,建立了CK-MB荧光免疫层析检测方法。荧光微球偶联率91.7%,CK-MB检测卡定量检测线性范围为1ng/ml~1000ng/ml,灵敏度达到了1ng/ml,CV值<8%,回收率84%~108.3%。利用间接ELISA法对市售抗体进行筛选,获得了包被抗体以及标记抗体,建立了心肌损伤标志物肌酸激酶同工酶荧光检测方法,实现血清中CK-MB定性与定量检测,为心肌损伤标志物荧光免疫层析检测方法的研发提供了实验技术基础。更多还原

【Abstract】 Objective:creatine kinase isoenzyme(CK-MB) is the early diagnosis of acute myocardial infarction(AMI) is an important marker of the fluorescent microspheres as a marker of rapid and sensitive fluorescence immunochromatographic detection method CK-MB. Methods:the fluorescent microspheres were covalently conjugated to commercially available antibodies by N-hydroxysuccinyl(NHS) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride(EDC),Optimizd the coupling conditions between antibody and fluorescent microspheres,characterized the complexes,prepared the fluorescent immunochromatographic assay card for CK-MB,estabilished the CK-MB fluorescence immunochromatographic assay. Results:coupling rate was 91.7%,detection linear range was 1 ng/ml~1000 ng/ml,sensitivity was 1 ng/ml,CV<8%,recovery rate was 84%~108.3%. Conclusion:the commercially available antibodies were screened by indirect ELISA method,obtained the coated antibody and the labeled antibody,proposed on creatine kinase isoenzyme fluorescence detection of myocardial injury markers,to achieve qualitative and quantitative detection of serum CK-MB,This study provides an experimental basis for the development of fluorescence immunochromatographic assays for myocardial injury markers.更多还原