一种灵敏的干血斑葡萄糖-6-磷酸脱氢酶荧光分析法的建立  

Development of A Sensitive and Rapid Fluorometric Method for the Measurement of Glucose-6-Phosphate Dehydrogenase in Dried Blood Spot Specimens

作　　者：汤永平[1] 李明[1] 徐伟文[1] 唐宁 黄晓春[3] 周才[4] 刘惠群 吴英松[1] 

机构地区：[1]南方医科大学生物技术学院,广州510515 [2]柳州市妇幼保健院,柳州545001 [3]深圳市妇幼保健院,深圳518028 [4]广东省妇幼保健院,广州510010 [5]广州市达瑞抗体工程技术有限公司,广州510665

出　　处：《热带医学杂志》2009年第11期1238-1240,1255,共4页Journal of Tropical Medicine

基　　金：广州市重大科技攻关项目(No.2005Z1-E4031)

摘　　要：目的建立葡萄糖-6-磷酸盐脱氢酶(G6PD)荧光分析方法,用于新生儿足跟干血斑中G6PD酶活性的定量测定。方法选择干血斑配制基质,优化底物、复溶物和铜试剂组分,建立G6PD荧光分析法定量检测方法,并对该试剂的各项性能指标进行评价。结果自制试剂分析灵敏度为0.05U/gHb;分析内和分析间精密度分别为4.12%～8.93%、4.75%～9.86%;试剂的cutoff值为2.23U/gHb;1066份样本用该试剂与PerkinElmer(PE)公司同类试剂平行检测,两种试剂测值具有显著相关性(P=0.000),相关系数为0.960;以PE公司试剂为对照,本试剂的阳性符合率为99.4%,阴性符合率为98.93%。结论该试剂灵敏度高、操作简便快捷,具有良好的精密度,临床检测性能良好,可以满足临床应用的需要。Objective To develop a quantitative fluorometric method for the detection of glucose-6-phosphate dehydrogenase （G6PD） in dried blood spot. Methods Reconstitution buffers and copper reagents were optimized for the testing of dried blood spot samples. The performance of the kit was then evaluated by testing 1 066 dried blood spot specimens. Results The assay sensitivity was found to be 0.05 U/g Hb. The intra- and inter-assay coefficients of variation were 4.12%-8.93% and 4.75%-9.86%, respectively. The cutoff value of the kit was 2.30 U/g Hb. Compared with the commercially available G6PD kit （PerkinElmer）, the positive coincidence rate of the present method was 99.4% and the negative coincidence rate was 98.93%. In addition, the two methods showed an excellent correlation, with a correlation coefficient of 0.960 （P=0.000）. Conclusion The developed G6PD kit is suitable for clinical application with better detection eapability.

关 键 词：葡萄糖-6-磷酸盐脱氢酶 荧光分析法 干血斑 

分 类 号：R446.61[医药卫生—诊断学]