Molecular combing compared to Southern blot for measuring D4Z4 contractions in FSHD
| Autor: | Khalida Liaquat, Malgorzata Jaremko, Aaron Bensimon, Patricia Chan, Carol Hoffman, Charles M. Strom, Sat Dev Batish, Arturo Anguiano, Marc Meservey, Renius Owen, Weimin Sun, Michael Jocson, Pierre Walrafen, Isabell Chang, Joseph Higgins, Jun Komatsu, Jessica Vasale, Jia-Chi Wang, Kerri Hensley, Yan Liu, Fatih Z Boyar, Zhenyuan Wang, David Tsao, Vladimira Sulcova, Corey D. Braastad |
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| Rok vydání: | 2015 |
| Předmět: |
animal structures
fungi Locus (genetics) Combing Sequence Analysis DNA Biology medicine.disease Molecular biology Muscular Dystrophy Facioscapulohumeral Blot genomic DNA Blotting Southern Neurology Gene mapping Molecular Diagnostic Techniques Pediatrics Perinatology and Child Health medicine Facioscapulohumeral muscular dystrophy Humans Neurology (clinical) Allele Chromosomes Human Pair 4 Genetics (clinical) Southern blot |
| Zdroj: | Neuromuscular disorders : NMD. 25(12) |
| ISSN: | 1873-2364 |
| Popis: | We compare molecular combing to Southern blot in the analysis of the facioscapulohumeral muscular dystrophy type 1 locus (FSHD1) on chromosome 4q35-qter (chr 4q) in genomic DNA specimens sent to a clinical laboratory for FSHD testing. A de-identified set of 87 genomic DNA specimens determined by Southern blot as normal (n = 71), abnormal with D4Z4 macrosatellite repeat array contractions (n = 7), indeterminate (n = 6), borderline (n = 2), or mosaic (n = 1) was independently re-analyzed by molecular combing in a blinded fashion. The molecular combing results were identical to the Southern blot results in 75 (86%) of cases. All contractions (n = 7) and mosaics (n = 1) detected by Southern blot were confirmed by molecular combing. Of the 71 samples with normal Southern blot results, 67 (94%) had concordant molecular combing results. The four discrepancies were either mosaic (n = 2), rearranged (n = 1), or borderline by molecular combing (n = 1). All indeterminate Southern blot results (n = 6) were resolved by molecular combing as either normal (n = 4), borderline (n = 1), or rearranged (n = 1). The two borderline Southern blot results showed a D4Z4 contraction on the chr 4qA allele and a normal result by molecular combing. Molecular combing overcomes a number of technical limitations of Southern blot by providing direct visualization of D4Z4 macrosatellite repeat arrays on specific chr 4q and chr 10q alleles and more precise D4Z4 repeat sizing. This study suggests that molecular combing has superior analytical validity compared to Southern blot for determining D4Z4 contraction size, detecting mosaicism, and resolving borderline and indeterminate Southern blot results. Further studies are needed to establish the clinical validity and diagnostic accuracy of these findings in FSHD. |
| Databáze: | OpenAIRE |
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