Simultaneous testing for 6 lysosomal storage disorders and x-adrenoleukodystrophy in dried blood spots by tandem mass spectrometry

Silvia Tortorelli, Coleman T. Turgeon, Dimitar K. Gavrilov, Devin Oglesbee, Kimiyo M. Raymond, Piero Rinaldo, Dietrich Matern

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

BACKGROUND: Newborn screening for lysosomal storage disorders (LSD) has revealed that late-onset variants of these conditions are unexpectedly frequent and therefore may evade diagnosis. We developed an efficient and costeffective multiplex assay to diagnose six LSDs and several peroxisomal disorders in patients presenting with diverse phenotypes at any age. METHODS: Three 3-mm dried blood spot (DBS) punches were placed into individual microtiter plates. One disc was treated with a cocktail containing acid sphingomyelinase-specific substrate and internal standard (IS). To the second DBS we added a cocktail containing substrate and IS for -glucosidase, acid -glucosidase, -galactosidase A, galactocerebrosidase, and -L-iduronidase. The third DBS was extracted with methanol containing d4-C26 lysophosphatidylcholine as IS and stored until the enzyme plates were combined and purified by liquid-liquid and solid-phase extraction. The extracts were evaporated, reconstituted with the extract from the lysophosphatidylcholine plate, and analyzed by flow injection tandem mass spectrometry. RESULTS: Reference intervals were determined by analysis of 550 samples from healthy controls. DBS from confirmed patients with 1 of the 6 LSDs (n 33), X-adrenoleukodystrophy (n 9), or a peroxisomal biogenesis disorder (n 5), as well as carriers for Fabry disease (n 17) and X-adrenoleukodystrophy (n 5), were analyzed for assay validation. Prospective clinical testing of 578 samples revealed 25 patients affected with 1 of the detectable conditions. CONCLUSIONS: Our flow injection tandem mass spectrometry approach is amenable to high-throughput population screening for Hurler disease, Gaucher disease, Niemann-Pick A/B disease, Pompe disease, Krabbe disease, Fabry disease, X-adrenoleukodystrophy, and peroxisomal biogenesis disorder in DBS.

Original languageEnglish (US)
Pages (from-to)1248-1254
Number of pages7
JournalClinical Chemistry
Volume62
Issue number9
DOIs
StatePublished - Sep 1 2016

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Adrenoleukodystrophy
Tandem Mass Spectrometry
Peroxisomal Disorders
Mass spectrometry
Blood
Glucosidases
Testing
Fabry Disease
Lysergic Acid Diethylamide
Lysophosphatidylcholines
Galactosylceramidase
Iduronidase
Globoid Cell Leukodystrophy
Galactosidases
Glycogen Storage Disease Type II
Mucopolysaccharidosis I
Sphingomyelin Phosphodiesterase
Gaucher Disease
Liquid-Liquid Extraction
Injections

ASJC Scopus subject areas

  • Medicine(all)
  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Simultaneous testing for 6 lysosomal storage disorders and x-adrenoleukodystrophy in dried blood spots by tandem mass spectrometry. / Tortorelli, Silvia; Turgeon, Coleman T.; Gavrilov, Dimitar K.; Oglesbee, Devin; Raymond, Kimiyo M.; Rinaldo, Piero; Matern, Dietrich.

In: Clinical Chemistry, Vol. 62, No. 9, 01.09.2016, p. 1248-1254.

Research output: Contribution to journalArticle

Tortorelli, Silvia ; Turgeon, Coleman T. ; Gavrilov, Dimitar K. ; Oglesbee, Devin ; Raymond, Kimiyo M. ; Rinaldo, Piero ; Matern, Dietrich. / Simultaneous testing for 6 lysosomal storage disorders and x-adrenoleukodystrophy in dried blood spots by tandem mass spectrometry. In: Clinical Chemistry. 2016 ; Vol. 62, No. 9. pp. 1248-1254.
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abstract = "BACKGROUND: Newborn screening for lysosomal storage disorders (LSD) has revealed that late-onset variants of these conditions are unexpectedly frequent and therefore may evade diagnosis. We developed an efficient and costeffective multiplex assay to diagnose six LSDs and several peroxisomal disorders in patients presenting with diverse phenotypes at any age. METHODS: Three 3-mm dried blood spot (DBS) punches were placed into individual microtiter plates. One disc was treated with a cocktail containing acid sphingomyelinase-specific substrate and internal standard (IS). To the second DBS we added a cocktail containing substrate and IS for -glucosidase, acid -glucosidase, -galactosidase A, galactocerebrosidase, and -L-iduronidase. The third DBS was extracted with methanol containing d4-C26 lysophosphatidylcholine as IS and stored until the enzyme plates were combined and purified by liquid-liquid and solid-phase extraction. The extracts were evaporated, reconstituted with the extract from the lysophosphatidylcholine plate, and analyzed by flow injection tandem mass spectrometry. RESULTS: Reference intervals were determined by analysis of 550 samples from healthy controls. DBS from confirmed patients with 1 of the 6 LSDs (n 33), X-adrenoleukodystrophy (n 9), or a peroxisomal biogenesis disorder (n 5), as well as carriers for Fabry disease (n 17) and X-adrenoleukodystrophy (n 5), were analyzed for assay validation. Prospective clinical testing of 578 samples revealed 25 patients affected with 1 of the detectable conditions. CONCLUSIONS: Our flow injection tandem mass spectrometry approach is amenable to high-throughput population screening for Hurler disease, Gaucher disease, Niemann-Pick A/B disease, Pompe disease, Krabbe disease, Fabry disease, X-adrenoleukodystrophy, and peroxisomal biogenesis disorder in DBS.",
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T1 - Simultaneous testing for 6 lysosomal storage disorders and x-adrenoleukodystrophy in dried blood spots by tandem mass spectrometry

AU - Tortorelli, Silvia

AU - Turgeon, Coleman T.

AU - Gavrilov, Dimitar K.

AU - Oglesbee, Devin

AU - Raymond, Kimiyo M.

AU - Rinaldo, Piero

AU - Matern, Dietrich

PY - 2016/9/1

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N2 - BACKGROUND: Newborn screening for lysosomal storage disorders (LSD) has revealed that late-onset variants of these conditions are unexpectedly frequent and therefore may evade diagnosis. We developed an efficient and costeffective multiplex assay to diagnose six LSDs and several peroxisomal disorders in patients presenting with diverse phenotypes at any age. METHODS: Three 3-mm dried blood spot (DBS) punches were placed into individual microtiter plates. One disc was treated with a cocktail containing acid sphingomyelinase-specific substrate and internal standard (IS). To the second DBS we added a cocktail containing substrate and IS for -glucosidase, acid -glucosidase, -galactosidase A, galactocerebrosidase, and -L-iduronidase. The third DBS was extracted with methanol containing d4-C26 lysophosphatidylcholine as IS and stored until the enzyme plates were combined and purified by liquid-liquid and solid-phase extraction. The extracts were evaporated, reconstituted with the extract from the lysophosphatidylcholine plate, and analyzed by flow injection tandem mass spectrometry. RESULTS: Reference intervals were determined by analysis of 550 samples from healthy controls. DBS from confirmed patients with 1 of the 6 LSDs (n 33), X-adrenoleukodystrophy (n 9), or a peroxisomal biogenesis disorder (n 5), as well as carriers for Fabry disease (n 17) and X-adrenoleukodystrophy (n 5), were analyzed for assay validation. Prospective clinical testing of 578 samples revealed 25 patients affected with 1 of the detectable conditions. CONCLUSIONS: Our flow injection tandem mass spectrometry approach is amenable to high-throughput population screening for Hurler disease, Gaucher disease, Niemann-Pick A/B disease, Pompe disease, Krabbe disease, Fabry disease, X-adrenoleukodystrophy, and peroxisomal biogenesis disorder in DBS.

AB - BACKGROUND: Newborn screening for lysosomal storage disorders (LSD) has revealed that late-onset variants of these conditions are unexpectedly frequent and therefore may evade diagnosis. We developed an efficient and costeffective multiplex assay to diagnose six LSDs and several peroxisomal disorders in patients presenting with diverse phenotypes at any age. METHODS: Three 3-mm dried blood spot (DBS) punches were placed into individual microtiter plates. One disc was treated with a cocktail containing acid sphingomyelinase-specific substrate and internal standard (IS). To the second DBS we added a cocktail containing substrate and IS for -glucosidase, acid -glucosidase, -galactosidase A, galactocerebrosidase, and -L-iduronidase. The third DBS was extracted with methanol containing d4-C26 lysophosphatidylcholine as IS and stored until the enzyme plates were combined and purified by liquid-liquid and solid-phase extraction. The extracts were evaporated, reconstituted with the extract from the lysophosphatidylcholine plate, and analyzed by flow injection tandem mass spectrometry. RESULTS: Reference intervals were determined by analysis of 550 samples from healthy controls. DBS from confirmed patients with 1 of the 6 LSDs (n 33), X-adrenoleukodystrophy (n 9), or a peroxisomal biogenesis disorder (n 5), as well as carriers for Fabry disease (n 17) and X-adrenoleukodystrophy (n 5), were analyzed for assay validation. Prospective clinical testing of 578 samples revealed 25 patients affected with 1 of the detectable conditions. CONCLUSIONS: Our flow injection tandem mass spectrometry approach is amenable to high-throughput population screening for Hurler disease, Gaucher disease, Niemann-Pick A/B disease, Pompe disease, Krabbe disease, Fabry disease, X-adrenoleukodystrophy, and peroxisomal biogenesis disorder in DBS.

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