精浆α葡糖苷酶全自动检测方法的建立及评价

Establishment and evaluation of an automatic method for seminal plasma α-glucosidase detection

目的：建立精浆α葡糖苷酶活性全自动检测方法并对其准确性、重复性和线性范围等进行评价。方法：利用速率法检测精浆α葡糖苷酶活性,并在全自动生化分析仪上建立检测参数,并评估该方法的试剂空白吸光度、准确性、重复性和线性范围。结果：试剂空白吸光度平均为0.002 14,空白吸光度变化率（△A/min）平均为0.015 26。3份高、中、低α葡糖苷酶活性的精浆样本重复测定10次的CV值分别为2.52%、0.36%和0.76%。采用比对试验的方法来评价准确性,40份精浆标本每个浓度点的相对偏差范围为-5.52%~7.54%,均低于15%的要求。精浆α葡糖苷酶活性在76~647 U/L范围时,具有良好的线性关系（r〉0.99）。结论：本研究建立的全自动精浆α葡糖苷酶活性检测法有较低的试剂空白,重复性和准确性较好,且有较宽的线性范围。该法操作简单、快速,精浆样本无需稀释,适合临床上大批量样本的检测筛查,大大节省了人力和试剂成本。

Objective： To establish an automatic method for seminal plasma α-glucosidase detection and evaluate its accuracy,repeatability,and linear range. Methods： We detected the α-glucosidase activity in the seminal plasma by rate assay,established the detection parameters on an automatic biochemical analyzer,and then evaluated the reagent blank absorbance,accuracy,repeatability,and linear range of the automatic method. Results： The average absorbance of reagent blank was 0. 002 14 and the mean change rate of blank absorbance（ △A /min） was 0. 015 26. The coefficients of variation（ CV） for 3 seminal plasma samples with high,middle and low α-glucosidase activity detected for 10 times,respectively,were 2. 52%,0. 36% and 0. 76%. With the accuracy of the automatic method evaluated by a comparison test,the relative deviation for each concentration point of 40 seminal plasma samples ranged from- 5. 52% to 7. 54%,which met the requirement of 15%. There was a good linear relationship（ r〉 0. 99） when the seminal plasma α-glucosidase activity was between 76 and 647 U /L. Conclusion： The established automatic method to detect α-glucosidase activity in the seminal plasma has a low reagent blank,good repeatability and accuracy,and a wide linear range. Simple,rapid,needing no dilution of the seminal plasma sample,and economical of manpower and reagents,it is suitable for screening large quantities of samples clinically.