title: Short-incubation mass spectrometry assay for lysosomal storage disorders in newborn and high-risk population screening.
abstract: The interest in early detection strategies for lysosomal storage disorders (LSDs) in newborns and high-risk population has increased in the last years due to the availability of novel treatment strategies coupled with the development of diagnostic techniques. We report the development of a short-incubation mass spectrometry-based protocol that allows the detection of Gaucher, Niemann-Pick A/B, Pompe, Fabry and mucopolysaccharidosis type I disease within 4h including sample preparation from dried blood spots. Optimized sample handling without the need of time-consuming offline preparations, such as liquid-liquid and solid-phase extraction, allows the simultaneous quantification of five lysosomal enzyme activities using a cassette of substrates and deuterated internal standards. Applying incubation times of 3h revealed in intra-day CV% values ranging from 4% to 11% for all five enzyme activities, respectively. In a first clinical evaluation, we tested 825 unaffected newborns and 16 patients with LSDs using a multiplexed, turbulent flow chromatography-ultra high performance liquid chromatography-tandem mass spectrometer assay. All affected patients were identified accurately and could be differentiated from non-affected newborns. In comparison to previously published two-day assays, which included an overnight incubation, this protocol enabled the detection of lysosomal enzyme activities from sample to first result within half a day.
authors.affiliation: Department of Pediatrics and Adolescent Medicine, Medical University of Vienna, Währinger Gürtel 18-20, Vienna, Austria.
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Mechtler
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Metz
authors.forename: Hannes G
authors.initials: HG
authors.lastname: Müller
authors.forename: Katharina
authors.initials: K
authors.lastname: Ostermann
authors.forename: Rene
authors.initials: R
authors.lastname: Ratschmann
authors.forename: Victor R
authors.initials: VR
authors.lastname: De Jesus
authors.forename: Bori
authors.initials: B
authors.lastname: Shushan
authors.forename: Joseph M
authors.initials: JM
authors.lastname: Di Bussolo
authors.forename: Joseph L
authors.initials: JL
authors.lastname: Herman
authors.forename: Kurt R
authors.initials: KR
authors.lastname: Herkner
authors.forename: David C
authors.initials: DC
authors.lastname: Kasper
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D059788:Dried Blood Spot Testing / Q000379:methods; D004355:Drug Stability; D057075:Enzyme Assays / Q000379:methods; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D007231:Infant, Newborn; D059625:Liquid-Liquid Extraction; D016464:Lysosomal Storage Diseases / Q000097:blood / Q000175:diagnosis* / Q000201:enzymology; D015997:Neonatal Screening / Q000379:methods*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*