A rapid and accurate methylation‐sensitive high‐resolution melting analysis assay for the diagnosis of Prader Willi and Angelman patients
| Autor: | Hiago Azevedo Cintra, Igor Ribeiro Ferreira, Elenice Ferreira Bastos, Leonardo Henrique Ferreira Gomes, Wilton Darleans dos Santos Cunha, Leticia Guida, Zilton Vasconcelos, Letícia Lopes Cabral Guimarães da Fonseca, Juan C. Llerena |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
MS‐HRM Male medicine.medical_specialty congenital hereditary and neonatal diseases and abnormalities lcsh:QH426-470 Method Locus (genetics) 030105 genetics & heredity Biology high‐resolution melting Polymerase Chain Reaction High Resolution Melt snRNP Core Proteins Epigenesis Genetic 03 medical and health sciences Chromosome 15 Angelman syndrome Genetics medicine Humans Allele Molecular Biology Genetics (clinical) DNA Primers Chromosomes Human Pair 15 Cytogenetics nutritional and metabolic diseases Nuclear Proteins DNA Methylation medicine.disease Prader Willi syndrome Uniparental disomy nervous system diseases lcsh:Genetics 030104 developmental biology DNA methylation Female MS‐PCR Prader-Willi Syndrome |
| Zdroj: | Molecular Genetics & Genomic Medicine Molecular Genetics & Genomic Medicine, Vol 7, Iss 6, Pp n/a-n/a (2019) |
| ISSN: | 2324-9269 |
| Popis: | Background Prader Willi (PWS) and Angelman (AS) syndromes are rare genetic disorders characterized by deletions, uniparental disomy, and imprinting defects at chromosome 15. The loss of function of specific genes caused by genetic alterations in paternal allele causes PWS while the absence in maternal allele results AS. The laboratory diagnosis of PWS and AS is complex and demands molecular biology and cytogenetics techniques to identify the genetic mechanism related to the development of the disease. The DNA methylation analysis in chromosome 15 at the SNURF‐SNRPN locus through MS‐PCR confirms the diagnosis and distinguishes between PWS and AS. Our study aimed to establish the MS‐PCR technique associated with High‐Resolution Melting (MS‐HRM) in PWS and AS diagnostic with a single pair of primers. Methods We collected blood samples from 43 suspected patients to a cytogenetic and methylation analysis. The extracted DNA was treated with bisulfite to perform comparative methylation analysis. Results MS‐HRM and MS‐PCR agreed in 100% of cases, identifying 19(44%) PWS, 3(7%) AS, and 21(49%) Normal. FISH analysis detected four cases of PWS caused by deletions in chromosome 15. Conclusion The MS‐HRM showed good performance with a unique pair of primers, dispensing electrophoresis gel analysis, offering a quick and reproducible diagnostic. |
| Databáze: | OpenAIRE |
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