丙氨酸氨基转移酶干化学检测方法的建立及试纸条的制备

Development of dry chemistry method for alanine aminotransferase

目的建立一种以丙酮酸氧化酶法为反应原理的丙氨酸氨基转移酶(Alanine aminotransferase,ALT)干化学检测方法,并制备试纸条。方法试纸条由扩散层、底物层和显色层组成,包被有反应试剂,用反射式光度计测定ALT活性。对制备的试纸条进行各项验证。结果试纸条测定罗氏低、高值质控品的批内变异系数分别为7.4%和4.9%;线性范围为10～1 000 U/L;试纸条测定肝素锂抗凝全血及其血浆的结果差异无统计学意义(P>0.05);标本中胆红素≤257μmol/L,抗坏血酸≤50 mg/L,丙酮酸≤0.5 mmol/L时,ALT测定结果不受影响;建立的干化学法与国际临床化学联合会(IFCC)推荐的方法相关性良好(r=0.988 9);试纸条的95%置信区间参考范围为0～40 U/L。结论建立的干化学法反应迅速,结果准确,操作简便,适用于ALT的即时检验。

Objective To develop a dry chemistry method for alanine aminotransferase （ALT） based on pyruvate oxidase method and prepare the test strip. Methods The test strip consisted of a sample spreading layer, a substrate layer and a color de- velopment layer, which was coated with reaction agent for determination of ALT activity by reflectance photometer. The prepared test strip was verified. Results The intra-CV of the test strip for low （48. 7 U/L） and high （127 U/L） level quality controls were 7.4% and 4. 9% respectively. The linear range of the test strip was 10 - 1 000 U/L. No significant difference was observed between the determination results of heparinized blood and plasma by the test strip （P 〉 0. 05）. The determination results of ALT were uninflu- enced when the bilirubin content in sample was not more than 257 μmol/L, the ascorbic acid content was not more than 50 mg/ L, and the pyruvate content was not more than 0. 5 mmol/L. The developed dry chemistry method showed good relationship to the method recommended by International Federation of Clinical Chemistry （IFCC）（r = 0. 988 9）. The reference range of 95% confi- dence interval of the test strip was 0 - 40 U / L. Conclusion The developed dry chemistry method was rapid, accurate and simple, which was suitable for immediate determination of ALT.