两点法测定腺苷脱氨酶及临床初步应用

Kinctic assay of adcnosinc deaminasse and preliminary clinical application

应用Ｔｒｉｎｄｅｒ反应建立了两点法测定体液中腺苷脱氨酶。该方法应用方便，测定快速、灵敏、准确。对酶反应最佳条件进行了实验探讨，底物Ｋｍ为０．２６ｍｍｏｌ／Ｌ，最佳ｐＨ７．４，延滞期５分钟，５～１５分钟测得酶活性基本一致。批内ＣＶ２．０３％，批间ＣＶ３．６８％。５５例健康人血清腺苷脱氨酶（ＡＤＡ）２１．３９±４．１７Ｕ／Ｌ（±ｓ），３５例结核性胸腹水ＡＤＡ活性为６５．８３±１８．９２Ｕ／Ｌ（±ｓ）。

n this paper,we described kinetlc analytical method of adenosine deaminase(ADA) in body fluids by Trinder’s.This method is simple,rapid,sensitive and accurate. Optimum reaction conditions for the activity of ADA were investigated.The substrate Km was 0.26mMol / L The extinction coeffcient for hypoxanthine at 515nm are 323×10￣4/mol·cm. Optimum pH was 7.40.The delay time was 5.0 min with-run and between-run precision(CV)were 3.06% and 3.68%, respectively.The mean serum ADA activity of 55 healthy subjects was 21.63 ±4.40U / L, The ADA activity of 35 tuberculous pleuritis in pleural effusion was 65.83 ±18.92U/L.