Dystrophin Protein Quantification as a Duchenne Muscular Dystrophy Diagnostic Biomarker in Dried Blood Spots Using Multiple Reaction Monitoring Tandem Mass Spectrometry: A Preliminary Study
Duchenne muscular dystrophy (DMD) is an X-linked recessive disorder characterized by progressive muscle loss, leading to difficulties in movement. Mutations in the DMD gene that code for the protein dystrophin are responsible for the development of DMD disorder, where the synthesis of this protein i...
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| Format: | Article |
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MDPI AG
2022-06-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/27/12/3662 |
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| author | Refat M. Nimer Khalid M. Sumaily Arwa Almuslat Mai Abdel Jabar Essa M. Sabi Mohammad A. Al-Muhaizea Anas M. Abdel Rahman |
| author_facet | Refat M. Nimer Khalid M. Sumaily Arwa Almuslat Mai Abdel Jabar Essa M. Sabi Mohammad A. Al-Muhaizea Anas M. Abdel Rahman |
| author_sort | Refat M. Nimer |
| collection | DOAJ |
| description | Duchenne muscular dystrophy (DMD) is an X-linked recessive disorder characterized by progressive muscle loss, leading to difficulties in movement. Mutations in the DMD gene that code for the protein dystrophin are responsible for the development of DMD disorder, where the synthesis of this protein is completely halted. Therefore, circulating dystrophin protein could be a promising biomarker of DMD disease. Current methods for diagnosing DMD have sensitivity, specificity, and reproducibility limitations. Herein, a quantitative liquid chromatography–tandem spectrometry (LC–MS/MS) technique in multiple reaction monitoring (MRM) mode was designed and validated for accurate dystrophin protein measurement in a dried blood spot (DBS). The method was successfully validated on the basis of international guidelines regarding calibration curves, precision, and accuracy. In addition, patients and healthy controls were used to test the amount of dystrophin protein circulating in DBS samples as a potential biomarker for DMD disorders. DMD patients were found to have considerably lower levels than controls. To the best of our knowledge, this is the first study to report dystrophin levels in DBS through LC–MS/MS as a diagnostic marker for DMD to the proposed MRM method, providing a highly specific and sensitive approach to dystrophin quantification in a DBS that can be applied in DMD screening. |
| first_indexed | 2024-03-09T22:57:41Z |
| format | Article |
| id | doaj.art-fa09770229124e6997d22fbd7b7d7616 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-09T22:57:41Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-fa09770229124e6997d22fbd7b7d76162023-11-23T18:09:11ZengMDPI AGMolecules1420-30492022-06-012712366210.3390/molecules27123662Dystrophin Protein Quantification as a Duchenne Muscular Dystrophy Diagnostic Biomarker in Dried Blood Spots Using Multiple Reaction Monitoring Tandem Mass Spectrometry: A Preliminary StudyRefat M. Nimer0Khalid M. Sumaily1Arwa Almuslat2Mai Abdel Jabar3Essa M. Sabi4Mohammad A. Al-Muhaizea5Anas M. Abdel Rahman6Department of Medical Laboratory Sciences, Jordan University of Science and Technology, Irbid 22110, JordanClinical Biochemistry Unit, Pathology Department, College of Medicine, King Saud University, Riyadh 11461, Saudi ArabiaMetabolomics Section, Department of Clinical Genomics, Center for Genome Medicine, King Faisal Specialist Hospital and Research Center (KFSH-RC), Zahrawi Street, Al Maather, Riyadh 11211, Saudi ArabiaMetabolomics Section, Department of Clinical Genomics, Center for Genome Medicine, King Faisal Specialist Hospital and Research Center (KFSH-RC), Zahrawi Street, Al Maather, Riyadh 11211, Saudi ArabiaClinical Biochemistry Unit, Pathology Department, College of Medicine, King Saud University, Riyadh 11461, Saudi ArabiaDepartment of Neurosciences, King Faisal Specialist Hospital and Research Centre, Riyadh 11211, Saudi ArabiaMetabolomics Section, Department of Clinical Genomics, Center for Genome Medicine, King Faisal Specialist Hospital and Research Center (KFSH-RC), Zahrawi Street, Al Maather, Riyadh 11211, Saudi ArabiaDuchenne muscular dystrophy (DMD) is an X-linked recessive disorder characterized by progressive muscle loss, leading to difficulties in movement. Mutations in the DMD gene that code for the protein dystrophin are responsible for the development of DMD disorder, where the synthesis of this protein is completely halted. Therefore, circulating dystrophin protein could be a promising biomarker of DMD disease. Current methods for diagnosing DMD have sensitivity, specificity, and reproducibility limitations. Herein, a quantitative liquid chromatography–tandem spectrometry (LC–MS/MS) technique in multiple reaction monitoring (MRM) mode was designed and validated for accurate dystrophin protein measurement in a dried blood spot (DBS). The method was successfully validated on the basis of international guidelines regarding calibration curves, precision, and accuracy. In addition, patients and healthy controls were used to test the amount of dystrophin protein circulating in DBS samples as a potential biomarker for DMD disorders. DMD patients were found to have considerably lower levels than controls. To the best of our knowledge, this is the first study to report dystrophin levels in DBS through LC–MS/MS as a diagnostic marker for DMD to the proposed MRM method, providing a highly specific and sensitive approach to dystrophin quantification in a DBS that can be applied in DMD screening.https://www.mdpi.com/1420-3049/27/12/3662dystrophinDuchenne muscular dystrophy (DMD)diagnostic biomarkerliquid chromatography–tandem spectrometry (LC–MS/MS)multiple reaction monitoring (MRM)dried blood spot (DBS) |
| spellingShingle | Refat M. Nimer Khalid M. Sumaily Arwa Almuslat Mai Abdel Jabar Essa M. Sabi Mohammad A. Al-Muhaizea Anas M. Abdel Rahman Dystrophin Protein Quantification as a Duchenne Muscular Dystrophy Diagnostic Biomarker in Dried Blood Spots Using Multiple Reaction Monitoring Tandem Mass Spectrometry: A Preliminary Study Molecules dystrophin Duchenne muscular dystrophy (DMD) diagnostic biomarker liquid chromatography–tandem spectrometry (LC–MS/MS) multiple reaction monitoring (MRM) dried blood spot (DBS) |
| title | Dystrophin Protein Quantification as a Duchenne Muscular Dystrophy Diagnostic Biomarker in Dried Blood Spots Using Multiple Reaction Monitoring Tandem Mass Spectrometry: A Preliminary Study |
| title_full | Dystrophin Protein Quantification as a Duchenne Muscular Dystrophy Diagnostic Biomarker in Dried Blood Spots Using Multiple Reaction Monitoring Tandem Mass Spectrometry: A Preliminary Study |
| title_fullStr | Dystrophin Protein Quantification as a Duchenne Muscular Dystrophy Diagnostic Biomarker in Dried Blood Spots Using Multiple Reaction Monitoring Tandem Mass Spectrometry: A Preliminary Study |
| title_full_unstemmed | Dystrophin Protein Quantification as a Duchenne Muscular Dystrophy Diagnostic Biomarker in Dried Blood Spots Using Multiple Reaction Monitoring Tandem Mass Spectrometry: A Preliminary Study |
| title_short | Dystrophin Protein Quantification as a Duchenne Muscular Dystrophy Diagnostic Biomarker in Dried Blood Spots Using Multiple Reaction Monitoring Tandem Mass Spectrometry: A Preliminary Study |
| title_sort | dystrophin protein quantification as a duchenne muscular dystrophy diagnostic biomarker in dried blood spots using multiple reaction monitoring tandem mass spectrometry a preliminary study |
| topic | dystrophin Duchenne muscular dystrophy (DMD) diagnostic biomarker liquid chromatography–tandem spectrometry (LC–MS/MS) multiple reaction monitoring (MRM) dried blood spot (DBS) |
| url | https://www.mdpi.com/1420-3049/27/12/3662 |
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