Targeted screening of succinic semialdehyde dehydrogenase deficiency (SSADHD) employing an enzymatic assay for γ-hydroxybutyric acid (GHB) in biofluids
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Authors
Finochiaro, S
Vogel, Kara R.
Ainslie, Garrett R.
Gibson, K. Michael
Volken, Carmen
Andresen-Streichert, Hilke
Salomons, Gajja S.
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Publication date
2016
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01A - Journal article
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Molecular Genetics and Metabolism Reports
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Pages / Duration
81-89
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Elsevier
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Abstract
Hypothesis
An enzymatic assay for quantification of γ-hydroxybutyric acid (GHB) in biofluids can be employed for targeted screening of succinic semialdehyde dehydrogenase deficiency (SSADHD) in selected populations.
Rationale
We used a two-tiered study approach, in which the first study (proof of concept) examined 7 urine samples derived from patients with SSADHD and 5 controls, and the second study (feasibility study) examined a broader sample population of patients and controls, including plasma.
Objective
Split samples of urine and plasma (anonymized) were evaluated by enzymatic assay, gas chromatography alone (proof of concept) and gas chromatography–mass spectrometry, and the results compared.
Method
Multiple detection methods have been developed to detect GHB. We evaluated an enzymatic assay which employs recombinant GHB dehydrogenase coupled to NADH production, the latter quantified on a Cobas Integra 400 Plus. Results: In our proof of concept study, we analyzed 12 urine samples (5 controls, 7 SSADHD), and in the feasibility study we evaluated 33 urine samples (23 controls, 10 SSADHD) and 31 plasma samples (14 controls, 17 SSADHD). The enzymatic assay carried out on a routine clinical chemistry analyzer was robust, revealing excellent agreement with instrumental methods in urine (GC-FID: r = 0.997, p ≤ 0.001; GC–MS: r = 0.99, p ≤ 0.001); however, the assay slightly over-estimated GHB levels in plasma, especially those in which GHB levels were low. Conversely, correlations for the enzymatic assay with comparator methods for higher plasma GHB levels were excellent (GC–MS; r = 0.993, p ≤ 0.001).
Conclusion
We have evaluated the capacity of this enzymatic assay to identify patients with SSADHD via quantitation of GHB. The data suggests that the enzymatic assay may be a suitable screening method to detect SSADHD in selected populations using urine. In addition, the assay can be used in basic research the elucidate the mechanism of the underlying disease or monitor GHB- levels for the evaluation of drug candidates.
Synopsis
An enzymatic assay for GHB in biofluids was evaluated as a screening method for SSADHD and found to be reliable in urine, but in need of refinement for application to plasma.
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2214-4269
Language
English
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Yes
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Published
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Citation
WERNLI, Cédric, S FINOCHIARO, André BÜTTLER, Daniel GYGAX, Kara R. VOGEL, Garrett R. AINSLIE, K. Michael GIBSON, Carmen VOLKEN, Hilke ANDRESEN-STREICHERT, Gajja S. SALOMONS und Erwin E. JANSEN, 2016. Targeted screening of succinic semialdehyde dehydrogenase deficiency (SSADHD) employing an enzymatic assay for γ-hydroxybutyric acid (GHB) in biofluids. Molecular Genetics and Metabolism Reports. 2016. S. 81–89. DOI 10.1016/j.ymgmr.2016.07.009. Verfügbar unter: http://hdl.handle.net/11654/23643
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