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Useful second-tier tests in expanded newborn screening of isovaleric acidemia and methylmalonic aciduria

  • Newborn Screening
  • Published:
Journal of Inherited Metabolic Disease

Abstract

Common use of pivalate-generating antibiotics in newborns in Japan and low cutoff value of C5-acylcarnitine (C5) to detect mild forms of isovaleric acidemia (IVA) led to 1,065 positive results from IVA screening among 146,000 newborns tested by tandem mass spectrometry over the last 3 years. Using our method to determine isovalerylglycine (IVG) levels in dried blood spots (DBS) as a second-tier test with IVG cutoff value of 0.5 nmol/ml in DBS, one patient with severe IVA was identified, and no recall of the second DBS was needed. Retrospective analysis revealed that most patients with moderate to severe forms of IVA have decreased free-carnitine levels shortly after birth and higher levels of IVG than those of C5, which suggests that this method is useful in evaluating the severity of IVA. Another second-tier test, to measure methylmalonic acid (MMA) levels in DBS by gas chromatography/mass spectrometry (GC/MS), has been developed to overcome difficulties in screening methylmalonic aciduria (MMAU) and propionic acidemia. Methanol extract from DBS was dried and derivatized using N-methyl-N-(tert-butyldimethylsilyl)-trifluoroacetamide. GC/MS was performed using splitless injection, electron-impact ionization, and selected ion monitoring for data recording. MMAU patients had much higher DBS concentrations of MMA (24.2–321.9 nmol/ml) than control newborns (0.34 ± 0.11 nmol/ml). MMA measurement in DBS was thought to provide useful information about the severity of MMAU, as MMAU patients with high levels of MMA had decreased levels of free carnitine and mildly increased levels of propionylcarnitine.

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Abbreviations

C2:

Acetylcarnitine

C3:

Propionylcarnitine

C5:

Acylcarnitine with 5-carbon-atom acyl group

DBS:

Dried blood spot

GC/MS:

Gas chromatography-mass spectrometry

IVA:

Isovaleric acidemia

IVG:

Isovalerylglycine

LC:

Liquid chromatography

MS/MS:

Tandem mass spectrometry

MMA:

Methylmalonic acid

MMAU:

Methylmalonic aciduria

PA:

Propionic acidemia

PGA:

Pivalate-generating antibiotics

SIM:

Selected ion monitoring

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Acknowledgments

This study was supported in part by a Grant-in-Aid for Scientific Research (C), Japan Society for the Promotion of Science, and grants from The Ministry of Health, Labor, and Welfare of Japan. The authors are very grateful to Dr. Y. Kobayashi (Shimane University), Dr. Y. Okano (Osaka City University), Dr. T. Ohnishi (Tokushima Municipal Hospital), Dr. N. Ishige (Tokyo Health Service Association), Dr. E. Naito (Tokushima University), and Dr. Y. Nishida (Kurashiki Central Hospital) for their assistance in the retrospective study.

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Authors and Affiliations

  1. Department of Health Science, Faculty of Medical Sciences, University of Fukui, 23 Matsuoka-Shimoaizuki, Eiheiji-cho, Fukui, 910–1193, Japan

    Yosuke Shigematsu

  2. Department of Pediatrics, Faculty of Medical Sciences, University of Fukui, Fukui, Japan

    Ikue Hata

  3. Department of Pediatrics, Hiroshima University Graduate School of Biomedical Sciences, Hiroshima, Japan

    Go Tajima

Authors
  1. Yosuke Shigematsu
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  2. Ikue Hata
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  3. Go Tajima
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Corresponding author

Correspondence to Yosuke Shigematsu.

Additional information

Communicated by: Piero Rinaldo

References to electronic databases: Isovaleric acidemia: OMIM #243500, methylmalonyl-CoA mutase deficiency: OMIM #25100.

Competing interest: None declared.

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Shigematsu, Y., Hata, I. & Tajima, G. Useful second-tier tests in expanded newborn screening of isovaleric acidemia and methylmalonic aciduria. J Inherit Metab Dis 33 (Suppl 2), 283–288 (2010). https://doi.org/10.1007/s10545-010-9111-9

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  • DOI: https://doi.org/10.1007/s10545-010-9111-9

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