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The Rapid and Sensitive Quantitative Determination of Galactose by Combined Enzymatic and Colorimetric Method: Application in Neonatal Screening

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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.

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Acknowledgments

This research project was financially supported by Biochemistry & Metabolic Disorders Research Center, Department of Research & Technology, Golestan University of Medical Sciences, Gorgan, Iran.

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Authors and Affiliations

  1. Biochemistry and Metabolic Disorders Research Center, Golestan University of Medical Sciences, Gorgan, Iran

    Anvarsadat Kianmehr & Morteza Oladnabi

  2. Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Golestan University of Medical Sciences, Gorgan, Iran

    Anvarsadat Kianmehr

  3. Immunogenetic Research Center, Mazandaran University of Medical Sciences, Sari, Iran

    Abdolkarim Mahrooz

  4. Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran

    Javad Ansari

  5. Department of Medical Genetics, Faculty of Advanced Medical Technologies, Golestan University of Medical Sciences, Gorgan, Iran

    Morteza Oladnabi

  6. Department of Biology, Bandar Jask Branch, Islamic Azad University, Bandar Jask, Iran

    Hamid Shahbazmohammadi

  7. Enzyme Technology Laboratory, Genetic and Metabolism Research Group, Pasteur Institute of Iran, Tehran, Iran

    Hamid Shahbazmohammadi

Authors
  1. Anvarsadat Kianmehr
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  2. Abdolkarim Mahrooz
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  3. Javad Ansari
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  4. Morteza Oladnabi
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  5. Hamid Shahbazmohammadi
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Correspondence to Morteza Oladnabi or Hamid Shahbazmohammadi.

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Kianmehr, A., Mahrooz, A., Ansari, J. et al. The Rapid and Sensitive Quantitative Determination of Galactose by Combined Enzymatic and Colorimetric Method: Application in Neonatal Screening. Appl Biochem Biotechnol 179, 283–293 (2016). https://doi.org/10.1007/s12010-016-1993-z

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  • DOI: https://doi.org/10.1007/s12010-016-1993-z

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