应用ELISA和滴金免疫测定法检测血清肌红蛋白

Application of ELISA and Dot Immunogold Filtration Assay in the Determination of Serum Myoglobin

从人心肌提取肌红蛋白（ｍｙｏｇｌｏｂｉｎ，Ｍｂ）抗原，制备抗Ｍｂ血清和单克隆抗体，在此基础上建立了检测血清Ｍｂ的ＥＬＩＳＡ和滴金免疫测定法（ｄｏｔｉｍｍｕｎｏｇｏｌｄｆｉｌｔｒａｔｉｏｎａｓｓａｙ，ＤＩＧＦＡ）。ＥＬＩＳＡ法检测范围２５～８００ｎｇ／ｍｌ。批内ＣＶ＜５％，批间ＣＶ＜７％。健康男、女献血员各３２例和７４例老年人，血清Ｍｂ测定结果，参考值上限分别为５５．４０、５０．６７和６５．７５ｎｇ／ｍｌ。１５例急性心肌梗塞（ＡＭＩ）患者血清Ｍｂ含量均大于１３０ｎｇ／ｍｌ．滴金免疫测定法可在５ｍｉｎ内完成检测，Ｍｂ含量＞１００ｎｇ／ｍｌ为阳性。７例ＡＭＩ患者检测结果均为阳性。两种方法结合应用对ＡＭＩ的早期诊断及疗效观察极有价值。

Human myoglobin(Mb)extracted from human cardiac muscles was used to raise polyclonal and monoclonal antibodies against Mb.On this basis ELISA and dot immunogold filtration assay(DIGFA)for the determination of serum Mb have been developed.For the Mb ELISA the range of determination was 25～800 ng/ml,and the within-and between-run CVs were below 5% and 7% respectively.Based on the results of determination of serum Mb level of 32 cases each of male and female blood donors and 74 cases of healthy old age individuals,the reference upper limit normal value were 55.40,50.67 and 65.75ng/ml respectively.The serum Mb level of 15 oasses of acute myocardial infarction (AMI)was all above 130ng/ml.DIGFA could be completed within 5 minuts.Serum Mb concentration higher than 100 ng/ml showed positive result.The tests of serum samples of 7 AMI patients were all positive.It could be concluded that the ELISA and DIGFA were very useful in early diagnosis and monitoring of AMI.