人类血清酪氨酸酶测定方法建立和评估.docVIP

人类血清酪氨酸酶测定方法建立和评估 　　【摘要】目的:建立血清酪氨酸酶活性检测的方法，并进行方法学评价。方法:采用酶速率法检测人类血清酪氨酸酶活性，应用NCCLS评价方案对方法进行线性、精密度、灵敏度、干扰试验和生理差异等方面的评估。同时对青年男性、女性各20例和7例黄褐斑女性患者检测血清酪氨酸酶活性，比较血清酪氨酸酶活性在正常男、女性间的差异，同时对黄褐斑女性患者血清酪氨酸酶活性进行初步分析。结果:血清酪氨酸酶活性检测有较好的检测范围和良好的线性范围，总CV为5．01％，总胆红素 　　【关键词】酪氨酸酶活性 方法学评价 　　 　　Determination of serum human tyrosinase the establishment and assessment methods 　　Dai YuzhuSengXiongwenChen Zhiyong 　　【Abstract】Objective：To establish a method for determining serum tyrosinase activity，and to assess it．Methods：The serum tyrosinase activity was determined by the rate of emzyne reaction， using the NCCLS to assess the method，including linearity,accuracy,sensitivity， interferentialexamination and physical difference．Meanwhile，the serum tyrosinase activity was detected in 20 male youth．20 female youth and 7 female chloasma patients．The defference of serum tyrosinase activity was compared between the healthy male and female control groups，and used for primary analyse to serum tyrosinase activity of female patients．Results：The method had a wide detect range and a well lineage．and its total CV was 5．01％．Such that total bilirubin was less than 150μmol／L，Hb was less than 2．0μmol／L，TG was less than 9．6μmol／L and Vitamine C with cunring dosages cant affect the detection．Meanwhile．serum tyrosinase activity of female patients was higher than referrence range of healthy female contro1．Conclusion：The method can be used to detect serum tyrosinase activity,reliable and used in the full-automatic biochemistry analyse instrument with high efficiency and precise，reliable result． 　　【Key words】The activity of tyrosinaseMethodologlcal evaluation 　　 　　酪氨酸酶(EC1．14．18．1，Tyrosinase)又称多酚氧化酶，是结构复杂的多亚基的含铜氧化还原酶，广泛存在于微生物、动植物及人体中。酪氨酸酶具有独特的双重催化功能，是生物体内黑色素合成的关键酶，与人的衰老，昆虫的伤口愈合与发???，果蔬的褐变有密切关系。鉴于临床需要，通过对文献［1、2、3］的调阅及对方法学的研究，建立了对人血清酪氨酸酶活性的全自动生化分析仪检测方法及相应的试剂。这将有利于临床大批量检测的需要，同时提高了结果的可重复性和准确度。 　　 　　1材料与方法 　　1．1标本来源 　　1．1．1健康对照组：在我院体检的青年健康者，男性20人，平均年龄21．8岁，女性20人，平均年龄20．3岁。空腹采取静脉血，分离血清，-20℃保存待测。 　　1．1．2实验组：由皮肤科医生选