Mifflin T E, Hamilton M, Hubbard E, Kline M J, Bruns D E
Department of Pathology, University of Virginia Health Sciences Center, Charlottesville 22908.
Clin Chem. 1989 Jan;35(1):110-4.
We studied a method for measuring the pancreatic isoenzyme of amylase (EC 3.2.1.1) by use of a mouse monoclonal antibody against human salivary-type amylase (Clin Chem 1985;31:1283) coupled indirectly to particles of polyvinylidene fluoride via polyclonal goat anti-mouse immunoglobulin. These particles, in 200 microL of a suspension, could remove salivary amylase (activity 2200 U/L) from an equal volume of serum in 5 min. Measurement of amylase activity in the supernatant fluids from treated sera thus provided an assay of pancreatic amylase. Precision studies at three activity concentrations yielded within-run CVs of 1.6% to 1.7% (n = 25) and total CVs of 2.2% to 5.1% (20 days). Salivary amylase added to each of 10 sera was completely (99.8%, SD 1.6%) removed. The new method (y) showed the following regression statistics when compared with an electrophoretic method (x): slope = 0.989 (SD 0.019), intercept = -0.220% (SD 1.48%), SEE 4.0%, n = 51. Similar respective regression values were found for urine samples: slope = 0.934 (SD 0.053), intercept = 2.3 U/L (SD 3.2), SEE 8.4 U/L, n = 26. The following respective values were found when the new method (y) was compared with the previously described immunoprecipitation assay (x): slope = 1.02 (SD 0.02), intercept = 2.2% (SD 1.4%), SEE 3.3%, n = 23 sera. Reference intervals for pancreatic amylase activity in serum were established for three different substrates: maltotetraose, maltopentaose, and p-nitrophenylheptaoside.
我们研究了一种通过使用抗人唾液型淀粉酶的小鼠单克隆抗体（Clin Chem 1985;31:1283）来测量淀粉酶（EC 3.2.1.1）胰腺同工酶的方法，该抗体通过山羊抗小鼠免疫球蛋白多克隆抗体间接偶联到聚偏二氟乙烯颗粒上。这些颗粒在200微升悬浮液中，5分钟内可从等体积血清中去除唾液淀粉酶（活性2200 U/L）。因此，测量处理后血清上清液中的淀粉酶活性可提供胰腺淀粉酶的测定方法。在三种活性浓度下进行的精密度研究得出批内变异系数为1.6%至1.7%（n = 25），总变异系数为2.2%至5.1%（20天）。添加到10份血清中的唾液淀粉酶被完全（99.8%，标准差1.6%）去除。与电泳法（x）相比，新方法（y）显示出以下回归统计数据：斜率 = 0.989（标准差0.019），截距 = -0.220%（标准差1.48%），估计标准误4.0%，n = 51。尿液样本也发现了类似的相应回归值：斜率 = 0.934（标准差0.053），截距 = 2.3 U/L（标准差3.2），估计标准误8.4 U/L，n = 26。当新方法（y）与先前描述的免疫沉淀法（x）相比时，得出以下相应值：斜率 = 1.02（标准差0.02），截距 = 2.2%（标准差1.4%），估计标准误3.3%，n = 23份血清。针对三种不同底物：麦芽四糖、麦芽五糖和对硝基苯基七糖苷，建立了血清中胰腺淀粉酶活性的参考区间。
