人肌型特异烯醇化酶放免分析法及初步应用

Radioimmunoassay of Human Muscle SpecificEnolase and Its Application.

建立了人肌型特异烯醇化酶（ｈＭＳＥ）的放免分析法，抗血清的亲和常数为５．１×１０￣９Ｌ／ｍｏｌ，采用改良的ＢＨＲ法制备了￣（１２５）Ｉ－ｈＭＳＥ，后者非特异结合率为３％，与抗血清（１∶１０￣３）结合率达５０．１６％；批内和批间ＣＶ分别为８．６％和１３％．回收率为９５％－１０５％．标准曲线范围为５－３２０μｇ／Ｌ．最小检出率为５μｇ／Ｌ．最佳反应条件：０．１ｍｏｌ／ＬｐＨ７．４ＰＢＳ（含５ｍｍｏｌ／ＬＭｇＳＯ＿４，０．１％吐温－２０，０．１％ＮａＮ＿３）；反应温度和时间：３７℃反应０．５ｈ和４℃，０．５ｈ，作为快速测定；或选用４℃反应２４ｈ．６５例健康人血清ｈＭＳＥ浓度为２３．９±１０．９μｇ／Ｌ（－ｘ±ｓ）．ｈＭＳＥ超过５７μｇ／Ｌ（－ｘ＋３ｓ）．为阳性值．测定了ＡＭＩ和肌病患者，血清ｈＭＳＥ明显升高．

Human muscle specific enolase(hMSE)plays an important role in the diag-nosis of acute myomuscle infarction and mus-cular diseases. A hMSE RIA is developed.Anaffinity coefficient of antiserum is 5.1×10￣9.￣(125)I-hMSE as the tracer is prepared using im-proved BHR method and its binding rate withantiserum(1∶1000)is up to 50.16%. Thevariable coefficients of intra and inter-assayare 8.6%and 13%, respectively. The recov-ery rate is 95%-105%。 The standards rangefrom 5 to 320μg/L and the minimum de-tectable value is 5μg/L. In the assay there isan optimal reaction condition:0.1mol/LpH7.4 PBS(5mmol/L MgSO_4,0.1%BSA,0.1%Toween-20,0.1% NaN_3,0.5%NaCl);and incabation temperature and times :37C℃,0.5h and 4℃,0.5h(as rapid measure-ment) or 4℃ 24h.On the basis of 65 healthycontrol,the hMSE levels are 23.9±10.9μg/L(x±s).A positive value is higher than57μg/L. The hMSE levels are signifcantly in-creased in the patients with both acute my-ocardial infarction and muscular diseases.