Tandem mass spectrometry assay of β-glucocerebrosidase activity in dried blood spots eliminates false positives detected in fluorescence assay

Pavlina Wolf; Roy N Alcalay; Christopher Liong; Emmaline Cullen; Michael W Pauciulo; William C Nichols; Ziv Gan-Or; Wendy K Chung; Tina Faulkner; Christopher Bentis; Robert J Pomponio; Xiwen Ma; X Kate Zhang; Joan M Keutzer; Petra Oliva

doi:10.1016/j.ymgme.2017.10.011

Tandem mass spectrometry assay of β-glucocerebrosidase activity in dried blood spots eliminates false positives detected in fluorescence assay

Mol Genet Metab. 2018 Feb;123(2):135-139. doi: 10.1016/j.ymgme.2017.10.011. Epub 2017 Oct 23.

Authors

Affiliations

¹ Sanofi, P. O. Box 9322, Framingham, MA 01701, USA.
² Columbia University Medical Center, Neurological Institute, 710 West, 168th street, New York, NY 10032, USA.
³ Sanofi, P. O. Box 9322, Framingham, MA 01701, USA; Great Ormond Street Hospital for Children, NHS Foundation Trust, Great Ormond Street, London, WC1N 3JH, UK.
⁴ Division of Human Genetics, Cincinnati Children's Hospital Medical Center and the Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH, USA.
⁵ Department of Neurology & Neurosurgery, Montreal Neurological Institute and Hospital, McGill University, Montreal, QC, Canada; Department of Human Genetics, McGill University, Montreal, QC, Canada.
⁶ Department of Pediatrics and Medicine, Columbia University Medical Center, New York, NY, USA.
⁷ Sanofi, P. O. Box 9322, Framingham, MA 01701, USA. Electronic address: petra.oliva@sanofi.com.

Abstract

Deficiency of β-Glucocerebrosidase (GBA) activity causes Gaucher Disease (GD). GD can be diagnosed by measuring GBA activity (Beutler and Kuhl, 1990). In this study, we assayed dried blood spots from a cohort (n=528) enriched for GBA mutation carriers (n=78) and GD patients (n=18) using both the tandem mass spectrometry (MS/MS) and fluorescence assays and their respective synthetic substrates. The MS/MS assay differentiated normal controls, which included GBA mutation carriers, from GD patients with no overlap. The fluorescence assay did not always differentiate normal controls including GBA mutation carriers from GD patients and false positives were observed. The MS/MS assay improved specificity compared to the fluorescence assay.

Keywords: Dried blood spots (DBS); Gaucher disease (GD); Glucocerebroside; Lysosomal storage disorder (LSD); Newborn screening (NBS); β-Glucocerebrosidase (GBA).

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Biological Assay
Biomarkers / blood*
Blood Specimen Collection
Case-Control Studies
Cohort Studies
Dried Blood Spot Testing*
Fluorescence*
Gaucher Disease / diagnosis*
Gaucher Disease / metabolism
Glucosylceramidase / blood*
Humans
Mass Screening*
Tandem Mass Spectrometry / methods*

Substances

Biomarkers
Glucosylceramidase

Abstract

Publication types

MeSH terms

Substances

Grants and funding