title: The Rapid and Sensitive Quantitative Determination of Galactose by Combined Enzymatic and Colorimetric Method: Application in Neonatal Screening.
abstract: The quantitative measurement of galactose in blood is essential for the early diagnosis, treatment, and dietary monitoring of galactosemia patients. In this communication, we aimed to develop a rapid, sensitive, and cost-effective combined method for galactose determination in dry blood spots. This procedure was based on the combination of enzymatic reactions of galactose dehydrogenase (GalDH), dihydrolipoyl dehydrogenase (DLD), and alkaline phosphates with a colorimetric system. The incubation time and the concentration of enzymes used in new method were also optimized. The analytical performance was studied by the precision, recovery, linearity, and sensitivity parameters. Statistical analysis was applied to method comparison experiment. The regression equation and correlation coefficient (R (2)) were Y = 0.0085x + 0.032 and R (2) = 0.998, respectively. This assay exhibited a recovery in the range of 91.7-114.3 % and had the limit detection of 0.5 mg/dl for galactose. The between-run coefficient of variation (CV) was between 2.6 and 11.1 %. The within-run CV was between 4.9 and 9.2 %. Our results indicated that the new and reference methods were in agreement because no significant biases exist between them. Briefly, a quick and reliable combined enzymatic and colorimetric assay was presented for application in newborn mass screening and monitoring of galactosemia patients.
authors.affiliation: Biochemistry and Metabolic Disorders Research Center, Golestan University of Medical Sciences, Gorgan, Iran.|Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Golestan University of Medical Sciences, Gorgan, Iran.
authors.forename: Anvarsadat
authors.identifier: http://orcid.org/0000-0001-5604-1870
authors.initials: A
authors.lastname: Kianmehr
authors.affiliation: Immunogenetic Research Center, Mazandaran University of Medical Sciences, Sari, Iran.
authors.forename: Abdolkarim
authors.initials: A
authors.lastname: Mahrooz
authors.affiliation: Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran.
authors.forename: Javad
authors.initials: J
authors.lastname: Ansari
authors.affiliation: Biochemistry and Metabolic Disorders Research Center, Golestan University of Medical Sciences, Gorgan, Iran. oladnabidozin@yahoo.com.|Department of Medical Genetics, Faculty of Advanced Medical Technologies, Golestan University of Medical Sciences, Gorgan, Iran. oladnabidozin@yahoo.com.
authors.forename: Morteza
authors.initials: M
authors.lastname: Oladnabi
authors.affiliation: Department of Biology, Bandar Jask Branch, Islamic Azad University, Bandar Jask, Iran. hamidbiozyme@gmail.com.|Enzyme Technology Laboratory, Genetic and Metabolism Research Group, Pasteur Institute of Iran, Tehran, Iran. hamidbiozyme@gmail.com.
authors.forename: Hamid
authors.initials: H
authors.lastname: Shahbazmohammadi
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D003124:Colorimetry / Q000379:methods; D004800:Enzymes, Immobilized / Q000737:chemistry; D005690:Galactose / Q000097:blood*; D005691:Galactose Dehydrogenases / Q000737:chemistry; D005693:Galactosemias / Q000097:blood* / Q000473:pathology; D006801:Humans; D007231:Infant, Newborn; D015997:Neonatal Screening
chemical_list: D004800:Enzymes, Immobilized; D005691:Galactose Dehydrogenases; C047959:galactose dehydrogenase; D005690:Galactose
keywords: Combined method; Galactose; Galactosemia; Newborn screening