Yamaguchi A, Fukushi M, Mizushima Y, Shimizu Y, Takasugi N, Arashima S, Ohyanagi K
Sapporo City Institute of Public Health, Japan.
Clin Chem. 1989 Sep;35(9):1962-4.
We describe a microassay for measuring galactose (Gal) and galactose 1-phosphate (Gal-1-P) in dried blood spots. After a coupled enzyme reaction involving galactose dehydrogenase (GADH, EC 1.1.1.48) and alkaline phosphatase (AP, EC 3.1.3.1) in a microplate well, NADH fluorescence is measured by a highly sensitive fluorometric microplate reader, capable of rapid measurement of fluorescence (2 min per 96 samples). Within- and between-run CVs for measurements of Gal at 90 mg/L with Gal-1-P at 130 mg/L were both less than 5% (n = 8), and analytical recoveries for Gal at 90 mg/L and Gal-1-P at 130 mg/L were 98% and 92%, respectively. Five hundred dried blood-spot samples can be assayed within 2 h, with full calculation of results by an on-line microcomputer. This rapid and reliable assay system is very useful for the routine screening of newborns for galactosemia.
我们描述了一种用于测量干血斑中半乳糖（Gal）和1-磷酸半乳糖（Gal-1-P）的微量测定法。在微孔板孔中进行涉及半乳糖脱氢酶（GADH，EC 1.1.1.48）和碱性磷酸酶（AP，EC 3.1.3.1）的偶联酶反应后，通过高灵敏度荧光微孔板读数仪测量NADH荧光，该读数仪能够快速测量荧光（每96个样品2分钟）。对于90 mg/L的Gal和130 mg/L的Gal-1-P测量，批内和批间变异系数均小于5%（n = 8），90 mg/L的Gal和130 mg/L的Gal-1-P的分析回收率分别为98%和92%。500个干血斑样品可在2小时内检测完毕，并通过在线微型计算机进行结果的完整计算。这种快速可靠的检测系统对于新生儿半乳糖血症的常规筛查非常有用。
