1. Using urine to diagnose large‐scale mtDNA deletions in adult patients
- Author
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Anu Suomalainen, Kristin N. Varhaug, Gonzalo S. Nido, Charalampos Tzoulis, Pirjo Isohanni, Irenaeus F.M. de Coo, Laurence A. Bindoff, Per M. Knappskog, RS: MHeNs - R3 - Neuroscience, Klinische Genetica, HUS Children and Adolescents, Research Programs Unit, Anu Wartiovaara / Principal Investigator, Clinicum, Children's Hospital, STEMM - Stem Cells and Metabolism Research Program, Faculty of Medicine, University of Helsinki, Helsinki University Hospital Area, HUSLAB, Department of Neurosciences, Neuroscience Center, and Helsinki Institute of Life Science HiLIFE
- Subjects
Male ,0301 basic medicine ,Ophthalmoplegia, Chronic Progressive External ,MITOCHONDRIAL-DNA ,Urine ,Polymerase Chain Reaction ,3124 Neurology and psychiatry ,law.invention ,0302 clinical medicine ,law ,Medicine ,Research Articles ,Polymerase chain reaction ,Sequence Deletion ,Sanger sequencing ,medicine.diagnostic_test ,General Neuroscience ,Mitochondrial Myopathies ,Middle Aged ,Heteroplasmy ,READ ALIGNMENT ,3. Good health ,Real-time polymerase chain reaction ,SINGLE ,symbols ,DETECT ,Female ,Research Article ,RC321-571 ,Adult ,Mitochondrial DNA ,Adolescent ,DISORDERS ,Neurosciences. Biological psychiatry. Neuropsychiatry ,PHENOTYPES ,Urinalysis ,DNA, Mitochondrial ,Sensitivity and Specificity ,DNA sequencing ,03 medical and health sciences ,symbols.namesake ,Humans ,RC346-429 ,Muscle biopsy ,business.industry ,3112 Neurosciences ,Sequence Analysis, DNA ,Molecular biology ,030104 developmental biology ,Neurology. Diseases of the nervous system ,Neurology (clinical) ,business ,030217 neurology & neurosurgery - Abstract
Objective: The aim of this study was to evaluate if urinary sediment cells offered a robust alternative to muscle biopsy for the diagnosis of single mtDNA deletions. Methods: Eleven adult patients with progressive external ophthalmoplegia and a known single mtDNA deletion were investigated. Urinary sediment cells were used to isolate DNA, which was then subjected to long‐range polymerase chain reaction. Where available, the patient`s muscle DNA was studied in parallel. Breakpoint and thus deletion size were identified using both Sanger sequencing and next generation sequencing. The level of heteroplasmy was determined using quantitative polymerase chain reaction. Results: We identified the deletion in urine in 9 of 11 cases giving a sensitivity of 80%. Breakpoints and deletion size were readily detectable in DNA extracted from urine. Mean heteroplasmy level in urine was 38% ± 26 (range 8 ‐ 84%), and 57% ± 28 (range 12 – 94%) in muscle. While the heteroplasmy level in urinary sediment cells differed from that in muscle, we did find a statistically significant correlation between these two levels (R = 0.714, P = 0.031(Pearson correlation)). Interpretation: Our findings suggest that urine can be used to screen patients suspected clinically of having a single mtDNA deletion. Based on our data, the use of urine could considerably reduce the need for muscle biopsy in this patient group. publishedVersion
- Published
- 2020