【摘要】 建立了测定腺苷脱氨酶（ＡＤＡ）的２，６二氯酚吲哚酚（ＤＣＩＰ）还原法，用腺苷（ＡＤ）作底物，通过核苷磷酸化酶（ＮＰ）和黄嘌呤氧化酶（ＸＯＤ）偶联反应，最终使蓝色ＤＣＩＰ还原成无色，在可见光波长下连续监测反应速率。ＤＣＩＰ的光吸收峰在６０２～６０４ｎｍ，为自动化需要，本法测定的波长为６００ｎｍ；ＤＣＩＰ的ε６００ｎｍ为１．７７４×１０４。每升底物缓冲液（ｐＨ７．２）中含１５７．５ｍｍｏｌ磷酸盐、１６．８ｍｍｏｌ腺苷（ＡＤ）、２００ＵＮＰ、２００ＵＸＯＤ和０．０７４ｍｍｏｌＤＣＩＰ。方法的批内ＣＶ为３．０％～３．３％、批间ＣＶ为５．９％～６．３％；样品中ＡＤＡ的活性在１８７．７Ｕ／Ｌ以内呈线性；ＮＨ３浓度在１５０ｍｍｏｌ／Ｌ以下不影响结果；参考范围为３０．５～４８．９Ｕ／Ｌ。更多还原

【Abstract】 The method for determination of adenosine deaminase(ADA)by means of the reduction of 2,6 dichlorophenolindophenol(DCIP)was established with adenosine(AD)as the substrate,nucleoside phosphorylase(NP)and xanthine oxidase(XOD)as coupling enzymes for the coupled reactions,in which blue DCIP is reduced to a colorless compound.ADA activity can be measured by detecting the reaction rate at visual wavelength continuously.Maximum absorbance of blue DCIP is at 602～604 nm.The wavelength of 600 nm was selected for the assay for automatic analyzer,at which DCIP has a mole absorptivity of 1 774×10 4.The substrate buffer contains phosphate of 157 5 mmol,AD of 16 8 mmol,NP of 200 U,XOD of 200 U and DCIP of 0 074 mmol per litre with pH 7 2.The obtained within run and between run CVs were of 3 0%～3 3% and 5 9%～6 3% respectively.The linearity range was up to 187 7 U/L of ADA in sample.NH 3 at the concentration less than 150 mmol/L does not interfer in the assay.The reference interval was obtained to be 30 5～48 9 U/L.更多还原