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目的对检测丙氨酸氨基转移酶(alanine aminotransferase,ALT)的双波长微板赖氏法进行优化。方法采用正交试验优化双波长微板赖氏法的空白试剂(磷酸缓冲液或纯化水)和Na OH浓度(0.5、1.0、1.5、2.0 mol/L),考察方法的稳定性、线性拟合度、精密性和准确性。结果以纯化水作空白,1.0 mol/L Na OH作终止溶液时,该方法具有很好的稳定性;线性拟合度好,R2=0.998 3>0.990 0;检测38和57卡门单位的丙酮酸标准溶液的板内A492/630值的变异系数均<10%,100%的样本检测结果在可接受范围内。结论优化的检测ALT的双波长微板赖氏法具有很好的稳定性、线性、精密性和准确性,且通量高,适用于大规模检测。
OBJECTIVE To optimize the dual-wavelength microplate assay for detecting alanine aminotransferase (ALT). Methods The blank reagents (phosphate buffer or purified water) and Na OH concentration (0.5, 1.0, 1.5 and 2.0 mol / L) were optimized by orthogonal test with two-wavelength microplate reader. The stability, Degree, precision and accuracy. The results showed that the method had good stability when using 1.0 mol / L NaOH as the stopping solution. The linearity was good, R2 = 0.998 3> 0.990 0. The pyruvate of 38 and 57 Carmen units The in-plate A492 / 630 values in the standard solutions all had a coefficient of variation of <10%, with 100% of the samples being within acceptable limits. Conclusion The optimized dual-wavelength microplate reader ALT has good stability, linearity, precision and accuracy, and high throughput, suitable for large-scale detection.