Targeted screening of succinic semialdehyde dehydrogenase deficiency (SSADHD) employing an enzymatic assay for γ-hydroxybutyric acid (GHB) in biofluids
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...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
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Elsevier
2017-06-01
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| Series: | Molecular Genetics and Metabolism Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S221442691630057X |
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| author | C. Wernli S. Finochiaro C. Volken H. Andresen-Streichert A. Buettler D. Gygax G.S. Salomons E.E. Jansen G.R. Ainslie K.R. Vogel K.M. Gibson |
| author_facet | C. Wernli S. Finochiaro C. Volken H. Andresen-Streichert A. Buettler D. Gygax G.S. Salomons E.E. Jansen G.R. Ainslie K.R. Vogel K.M. Gibson |
| author_sort | C. Wernli |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-21T20:06:22Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2214-4269 |
| language | English |
| last_indexed | 2024-12-21T20:06:22Z |
| publishDate | 2017-06-01 |
| publisher | Elsevier |
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| series | Molecular Genetics and Metabolism Reports |
| spelling | doaj.art-441db346b2a049eb8763cdb132d8cc992022-12-21T18:51:49ZengElsevierMolecular Genetics and Metabolism Reports2214-42692017-06-0111C818910.1016/j.ymgmr.2016.07.009Targeted screening of succinic semialdehyde dehydrogenase deficiency (SSADHD) employing an enzymatic assay for γ-hydroxybutyric acid (GHB) in biofluidsC. Wernli0S. Finochiaro1C. Volken2H. Andresen-Streichert3A. Buettler4D. Gygax5G.S. Salomons6E.E. Jansen7G.R. Ainslie8K.R. Vogel9K.M. Gibson10University of Applied Sciences and Arts, Northwestern Switzerland School of Life Sciences, Institute for Chemistry and Bioanalytics, SwitzerlandBildungszentrum Gesundheit Basel-Stadt, Münchenstein, SwitzerlandZentrallabor, Kantonsspital Baselland, Bruderholz, SwitzerlandUniversity Medical Center Hamburg-Eppendorf Institute of Legal Medicine, Department of Toxicology, Hamburg, GermanyUniversity of Applied Sciences and Arts Northwestern Switzerland School of Life Sciences, Institute for Chemistry and Bioanalytics Muttenz, SwitzerlandUniversity of Applied Sciences and Arts Northwestern Switzerland School of Life Sciences, Institute for Chemistry and Bioanalytics Muttenz, SwitzerlandMetabolic Unit, Department of Clinical Chemistry, VU University Mediscal Center, Neuroscience Campus, Amsterdam, The NetherlandsMetabolic Unit, Department of Clinical Chemistry, VU University Mediscal Center, Neuroscience Campus, Amsterdam, The NetherlandsDivision of Experimental and Systems Pharmacology, College of Pharmacy, Washington State University, Spokane, WA, USADivision of Experimental and Systems Pharmacology, College of Pharmacy, Washington State University, Spokane, WA, USADivision of Experimental and Systems Pharmacology, College of Pharmacy, Washington State University, Spokane, WA, USAHypothesis: 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.http://www.sciencedirect.com/science/article/pii/S221442691630057XSuccinic semialdehyde dehydrogenaseSuccinic semialdehyde dehydrogenase deficiencyγ-hydroxybutyric acidEnzymatic enzyme assay |
| spellingShingle | C. Wernli S. Finochiaro C. Volken H. Andresen-Streichert A. Buettler D. Gygax G.S. Salomons E.E. Jansen G.R. Ainslie K.R. Vogel K.M. Gibson Targeted screening of succinic semialdehyde dehydrogenase deficiency (SSADHD) employing an enzymatic assay for γ-hydroxybutyric acid (GHB) in biofluids Molecular Genetics and Metabolism Reports Succinic semialdehyde dehydrogenase Succinic semialdehyde dehydrogenase deficiency γ-hydroxybutyric acid Enzymatic enzyme assay |
| title | Targeted screening of succinic semialdehyde dehydrogenase deficiency (SSADHD) employing an enzymatic assay for γ-hydroxybutyric acid (GHB) in biofluids |
| title_full | Targeted screening of succinic semialdehyde dehydrogenase deficiency (SSADHD) employing an enzymatic assay for γ-hydroxybutyric acid (GHB) in biofluids |
| title_fullStr | Targeted screening of succinic semialdehyde dehydrogenase deficiency (SSADHD) employing an enzymatic assay for γ-hydroxybutyric acid (GHB) in biofluids |
| title_full_unstemmed | Targeted screening of succinic semialdehyde dehydrogenase deficiency (SSADHD) employing an enzymatic assay for γ-hydroxybutyric acid (GHB) in biofluids |
| title_short | Targeted screening of succinic semialdehyde dehydrogenase deficiency (SSADHD) employing an enzymatic assay for γ-hydroxybutyric acid (GHB) in biofluids |
| title_sort | targeted screening of succinic semialdehyde dehydrogenase deficiency ssadhd employing an enzymatic assay for γ hydroxybutyric acid ghb in biofluids |
| topic | Succinic semialdehyde dehydrogenase Succinic semialdehyde dehydrogenase deficiency γ-hydroxybutyric acid Enzymatic enzyme assay |
| url | http://www.sciencedirect.com/science/article/pii/S221442691630057X |
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