Development and Evaluation of a Hexokinase/Glucose-6-Phosphate Dehydrogenase Procedure for Use as a National Glucose Reference Method,

摘要

Results are presented of a comprehensive study in which over 10,000data pieces were generated to develop a national glucose reference method. A review of hexokinase (HK)/glucose-6-phosphate dehydrogenase (gpd) glucose methodology is included,and results of reaction parameter studies for four HK/GPD procedures are given. The approaches studied used yeast HK with either yeast GPD and NADP+as coenzyme or bacterial GPD (leuconostoc mesenteroids) and NAD+. Both protein precipitation and sample-in-saline blanking were investigated for these two enzyme systems. The resultant four HK/GPD methods, a manual glucose oxidase procedure,and an automated ferricyanide method were evaluated and compared for linearity of calibration curves. Precision,recovery of added glucose,response to 18possible interferring substances in standard and serum,and results for plasma specimens collected from 64normal and 92diabetic donors. The hexokinase/bacterial GPD/NAD+system applied to a Somogyi deproteinized supernatant was selected as the candidate glucose reference method,which was used for additional studies. The recommended procedure,along with materials and performance specifications,is given.