两点法测定腺苷脱氨酶

DETERMINATION OF ADENOSINE DEAMINASE BY TWO-POINT ASSAY

目的：基于文献报道法建立一种两点法测定腺苷脱氨酶（ＡＤＡ）活性的方法。方法：用２．４－二氯酚参与Ｔｒｉｎｄｅｒ反应建立了两点法，从标本用量和方法上进行改进，并进行方法学评价。结果：酶促反应时间短（１０分钟）；线性范围宽（０～１００Ｕ／Ｌ）；批内ＣＶ为２．２％～３．８％；日间ＣＶ为４．３％；Ｈｂ大于０．５６ｇ／Ｌ时呈正干扰，胆红素大于１７１．２μｍｏｌ／Ｌ时呈负干扰；１２７例健康人血清ＡＤＡ为２０．５±４．５７ｕ／Ｌ（ｘ±ｓ），２７例结核性胸腹水患者ＡＤＡ为７６．１５±１６．４Ｕ／Ｌ（ｘ±ｓ），而２１例非结核性胸腹水患者ＡＤＡ为２２．７０±１１．７０Ｕ／Ｌ，两组间有显著性差异。结论：本法线性范围宽，呈色稳定，重复性好，是结核性与非结核性胸腹水鉴别的特异指标。由于该法简便、快速、易于控制，可在教学单位及中小医院推广应用，具有一定价值。

Objective:To establish a two-point assay for measuring body fluid adenosine deaminase(ADA)activity based on reported methods.Methods:Using 2,4-phenyldichloride instead of TBHBA in Trinders reaction,a new two-point assay for measuring ADA activity was established.The sample sige and procedure were modified and methodological evaluation of the assay was carried out.Results:The reaction was shortened to 10 min;the linearity sange was wide (0～100U/L);the CVs of within batch and day to day were 2.2%～3.8%and 4.3% respectively;the Hb level above 0.56g/L gave a positive interference and the bilirubin level reaching 171.2μmol/L gave a negative interference;ADA activity for normal subjects ( n =127)is 20.5±4.57U/L,for pleurad and peritoncal effusions of tuberculous patients( n =27)is 76.15±16.4U/Land for non tuber culouspatients ( n =21)is 22.70±11.70U/L( P <0.05).Conclusion:The new assay showed wide linearity,good recovery and stable colouration and is a specific indication for distinguishing tuberculous effusions from non-tuberculous effusions.The method is simple,rapid and easily to control.It is worthy to be populariged in teaching and in application in middle-siged and small hospitals.