酶偶联放大法测定血清总胆汁酸

Determination of total bile acids in serum with an enzyme coupling amplification technique

目的 建立简便的酶偶联放大法 ,直接检测血清总胆汁酸 (TBA)。方法 基于脱氢酶 辅酶体系的原理 ,用丙酮酸钠作乳酸脱氢酶 (LDH)的阻止剂 ,在Tris HCl缓冲溶液中 ,由NAD+ -NBT -Brij 98组成酶偶联放大系统。用自动生化分析仪测定血清TBA。结果 线性范围 :0～ 180 μmol/L ;精密度 :批内CV <2 .7% ,日间CV <4 .3% ,总CV <5 .4 %。与日本ENZABILE .AUTO(X)比较 :r =0 .990 ,Y =1.114X - 2 .137,与朗道BILEACIDS(Z)比较 :r =0 .997,Y =1.0 32Z +0 .90 6。LDH <3kU/L ,乳酸 <2 0mmol/L ,维生素C <0 .5mmol/L ,胆红素 <30 0 μmol/L ,血红蛋白 <5g/L时 ,对结果无显著干扰。 结论 该法简便、灵敏 ,试剂稳定 。

Objective To establish a simple and direct method for determining total bile acids(TBA) in serum. Methods Based on the principle of dehydrogenase coenzyme system, sodium pyruvate was used as a lactate dehydrogenase(LDH) blocker, NAD +-NBT-Brij 98 was composed to enzyme coupling amplification technique in Tris HCl buffer. TBA was determined with a biochemical auto analyzer. Results The linearity was 0-180 μmol/L; The precision was within run CV <2.7%, between\|run CV <4.3%, and total CV <5.4%. Correlation between results obtained by this method(Y) and Japanese ENZABILE AUTO(X) was Y=1.114X-2.137, r =0.990 and RANDOX BILE ACIDS(Z) was Y=1.032Z+0.906, r =0.997. No interference was showed when LDH<3 kU/L, lactic acid<20 mmol/L, ascorbic acid<0.5 mmol/L, bilirubin<300 μmol/L, Hb<5 g/L were added to a sample in which TBA concentration was 108 μmol/L. Conclusions The method is simple sensitive and stable for routine and automatic measurement of TBA in serum.