Korean J Lab Med.  2011 Oct;31(4):250-256. 10.3343/kjlm.2011.31.4.250.

Use of Tandem Mass Spectrometry for Newborn Screening of 6 Lysosomal Storage Disorders in a Korean Population

• Han M
• Jun SH
• Song SH
• Park KU
• Kim JQ
• Song J

• KMID: 1781688
• DOI: http://doi.org/10.3343/kjlm.2011.31.4.250

Abstract

BACKGROUND
We evaluated the performance of multiplex tandem mass spectrometry (MS/MS) in newborn screening for detection of 6 lysosomal storage disorders (LSDs), namely, Niemann-Pick A/B, Krabbe, Gaucher, Fabry, and Pompe diseases and Hurler syndrome.
METHODS
We revised the conditions and procedures of multiplex enzyme assay for the MS/MS analysis and determined the precision of our enzyme assay and the effects of sample amounts and incubation time on the results. We also measured the degree of correlation between the enzyme activities in the dried blood spots (DBSs) and those in the leukocytes. DBSs of 211 normal newborns and 13 newborns with various LSDs were analyzed using our revised methods.
RESULTS
The intra- and inter-assay precisions were 2.9-18.7% and 8.1-18.1%, respectively. The amount of product obtained was proportional to the DBS eluate volume, but a slight flattening was observed in the product vs. sample volume curve at higher sample volumes. For each enzyme assay, the amount of product obtained increased linearly with the incubation period (range, 0-24 hr). Passing and Bablok regression analysis revealed that the enzyme activities in the DBSs and those in the leukocytes were favorably correlated. The enzyme activities measured in the DBSs were consistently lower in patients with LSDs than in normal newborns.
CONCLUSIONS
The performance of our revised techniques for MS/MS detection and enzyme assays was of the generally acceptable standard. To our knowledge, this is the first report on the use of MS/MS for newborn screening of LSDs in an Asian population.

Keyword

Lysosomal storage disorders; Multiplex enzyme assay; Tandem mass spectrometry; Newborn screening

MeSH Terms

Dried Blood Spot Testing
Enzyme Assays
Enzymes/blood
Humans
Infant, Newborn
Leukocytes/enzymology
Lysosomal Storage Diseases/*diagnosis
Republic of Korea
Tandem Mass Spectrometry/*methods
Time Factors

Figure

• 

  Fig. 1 Effect of sample amount on enzyme activity. Representative graphs for the 6 enzyme assays with respect to the added volume of DBS eluate.Abbreviations: ASM, acid sphingomyelinase (Niemann-Pick A/B disease); GALC, galactocerebrosidase (Krabbe disease); ABG, acid β-glucocerebrosidase (Gaucher disease); GLA, acid α-galactosidase (Fabry disease); GAA, acid α-glucosidase (Pompe disease); and IDU, α-L-iduronidase (Hurler disease).

• 

  Fig. 2 Effect of incubation time on enzyme activity. Representative graphs for the time course of the 6 enzyme assays. Each enzyme reaction was monitored for 24 hr.Abbreviations: ASM, acid sphingomyelinase (Niemann-Pick A/B disease); GALC, galactocerebrosidase (Krabbe disease); ABG, acid β-glucocerebrosidase (Gaucher disease); GLA, acid α-galactosidase (Fabry disease); GAA, acid α-glucosidase (Pompe disease); and IDU, α-L-iduronidase (Hurler disease).

• 

  Fig. 3 Passing and Bablok regression analysis for the lysosomal enzyme activities in the DBSs and leukocytes.Abbreviations: GALC, galactocerebrosidase (Krabbe disease); ABG, acid β-glucocerebrosidase (Gaucher disease); GLA, acid α-galactosidase (Fabry disease); GAA, acid α-glucosidase (Pompe disease); and IDU, α-L-iduronidase (Hurler disease).

• 

  Fig. 4 Comparison of enzyme activities in DBSs from healthy newborns (n=211) and patients with LSDs (4 patients with Krabbe disease, 1 patient with Gaucher disease, 1 patient with Fabry disease, 6 patients with Pompe disease, and 1 patient with Hurler syndrome).Abbreviations: ASM, acid sphingomyelinase (Niemann-Pick A/B disease); GALC, galactocerebrosidase (Krabbe disease); ABG, acid β-glucocerebrosidase (Gaucher disease); GLA, acid α-galactosidase (Fabry disease); GAA, acid α-glucosidase (Pompe disease); and IDU, α-L-iduronidase (Hurler disease).

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