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. 2019 Mar;5(1):1.
doi: 10.3390/ijns5010001. Epub 2018 Dec 21.

Newborn Screening for Lysosomal Storage Disorders: Methodologies for Measurement of Enzymatic Activities in Dried Blood Spots

Michael H Gelb  1 , Zoltan Lukacs  2 , Enzo Ranieri  3 , Peter C J I Schielen  4
Affiliations

Newborn Screening for Lysosomal Storage Disorders: Methodologies for Measurement of Enzymatic Activities in Dried Blood Spots

Michael H Gelb et al. Int J Neonatal Screen. 2019 Mar.

Abstract

All worldwide newborn screening (NBS) for lysosomal storage diseases (LSDs) is performed as a first-tier test by measurement of lysosomal enzymatic activities in dried blood spots (DBS). The currently two available methodologies used for measurement of enzymatic activities are tandem mass spectrometry (MS/MS) and digital microfluidics fluorimetry (DMF-F). In this chapter we summarize the workflows for the two platforms. Neither platform is fully automated, but the relative ease of workflow will be dependent upon the specific operation of each newborn screening laboratory on a case-by-case basis. We provide the screen positive rate (the number of below cutoff newborns per 100,000 newborns) from all NBS laboratories worldwide carrying out MS/MS-based NBS of one or more LSDs. The analytical precision of the MS/MS method is higher than that for DMF-F as shown by analysis of a common set of quality control DBS by the Centers for Disease Control and Prevention (CDC). Both the MS/MS and DMF-F platforms enable multiplexing of the LSD enzymes. An advantage of MS/MS over DMF-F is the ability to include assays of enzymatic activities and biomarkers for which no fluorimetric methods exist. Advantages of DMF-F over MS/MS are: 1) Simple to use technology with same-day turn-around time for the lysosomal enzymes with the fastest rates compared to MS/MS requiring overnight analytical runs.; 2) The DMF-F instrumentation, because of its simplicity, requires less maintenance than the MS/MS platform.

Keywords: cutoff values; diagnosis; dried blood spots; enzymatic activity assays; lysosomal storage diseases; newborn screening; tandem mass spectrometry.

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Conflict of interest statement

Gelb is a consultant for PerkinElmer and has received research funding from Biomarin, Shire, and Ultrageneyx corporations. Lukacs received travel and research grants and honoraria from Sanofi Genzyme B.V., Netherlands, BioMarin Ltd, Dublin, Ireland, Alexion Pharma GmbH, Munich, Germany, Ultragenyx, Zurich, Switzerland, PerkinElmer Life Science, Turku, Finland.

Figures

Figure 1
Figure 1
Plots of the precision data taken from the certificate reports for the Centers for Disease Control and Prevention (CDC) quality control DBS for GAA (Pompe disease) and alpha-iduronidase (IDUA, MPS-I) using either DMF-F or MS/MS (reports provided as Supplemental Material). The CDC carried out repetitive assays using either DMF-F or MS/MS with 10 punches from each quality control DBS. Plotted is the mean enzymatic activity (black dots) for the BASE sample (0 % QC level), the LOW sample (5% QC level), the MEDIUM sample (50% QC level), and the HIGH sample (100% QC level). The red dots are the 95% upper and lower confidence intervals.
See this image and copyright information in PMC

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