Methylation-sensitive high-resolution melting-curve analysis of the SNRPN gene as a diagnostic screen for Prader-Willi and Angelman syndromes
Methylation-sensitive high-resolution melting-curve analysis of the SNRPN gene as a diagnostic screen for Prader-Willi and Angelman syndromes
BACKGROUND: Angelman syndrome (AS) and Prader-Willi syndrome (PWS) are 2 distinct neurodevelopmental disorders caused primarily by deficiency of specific parental contributions at an imprinted domain within the chromosomal region 15q11.2-13. Lack of paternal contribution results in PWS either by paternal deletion (approximately 70%) or maternal uniparental disomy (UPD) (approximately 25%). Most cases of AS result from the lack of a maternal contribution from this same region, by maternal deletion (70%) or paternal UPD (approximately 5%). Analysis of allelic methylation differences at the small nuclear ribonucleoprotein polypeptide N (SNRPN) locus differentiates the maternally and paternally inherited chromosome 15 and can be used as a diagnostic test for AS and PWS. METHODS: Methylation-sensitive high-resolution melting-curve analysis (MS-HRM) using the DNA binding dye EvaGreen was used to analyze methylation differences at the SNRPN locus in anonymized DNA samples from individuals with PWS (n = 39) or AS (n = 31) and from healthy control individuals (n = 95). Results from the MS-HRM assay were compared to those obtained by use of a methylation-specific PCR (MSP) protocol that is used commonly in diagnostic practice. RESULTS: With the MS-HRM assay 97.6% of samples were unambiguously assigned to the 3 diagnostic categories (AS, PWS, normal) by use of automated calling with an 80% confidence percentage threshold, and the failure rate was 0.6%. One PWS sample showed a discordant result for the MS-HRM assay compared to MSP data. CONCLUSIONS: MS-HRM is a simple, rapid, and robust method for screening methylation differences at the SNRPN locus and could be used as a diagnostic screen for PWS and AS
chromosomes, syndrome, methods, deficiency, polymerase chain reaction, genotype, research support, genetics, uniparental disomy, diagnosis, angelman syndrome, ribonucleoproteins, autoantigens, human, pair 15, metabolism, analysis, transition temperature, prader-willi syndrome, antigens, dna methylation, laboratories, dna, research, small nuclear, humans
1960-1962
White, Helen E.
2181c0b9-fc3b-407e-95eb-3510524603e5
Hall, Victoria J.
a60eea2a-3fb9-460e-a845-692f8970e68a
Cross, Nicholas C.P.
f87650da-b908-4a34-b31b-d62c5f186fe4
1 November 2007
White, Helen E.
2181c0b9-fc3b-407e-95eb-3510524603e5
Hall, Victoria J.
a60eea2a-3fb9-460e-a845-692f8970e68a
Cross, Nicholas C.P.
f87650da-b908-4a34-b31b-d62c5f186fe4
White, Helen E., Hall, Victoria J. and Cross, Nicholas C.P.
(2007)
Methylation-sensitive high-resolution melting-curve analysis of the SNRPN gene as a diagnostic screen for Prader-Willi and Angelman syndromes.
Clinical Chemistry, 53 (11), .
(doi:10.1373/clinchem.2007.093351).
Abstract
BACKGROUND: Angelman syndrome (AS) and Prader-Willi syndrome (PWS) are 2 distinct neurodevelopmental disorders caused primarily by deficiency of specific parental contributions at an imprinted domain within the chromosomal region 15q11.2-13. Lack of paternal contribution results in PWS either by paternal deletion (approximately 70%) or maternal uniparental disomy (UPD) (approximately 25%). Most cases of AS result from the lack of a maternal contribution from this same region, by maternal deletion (70%) or paternal UPD (approximately 5%). Analysis of allelic methylation differences at the small nuclear ribonucleoprotein polypeptide N (SNRPN) locus differentiates the maternally and paternally inherited chromosome 15 and can be used as a diagnostic test for AS and PWS. METHODS: Methylation-sensitive high-resolution melting-curve analysis (MS-HRM) using the DNA binding dye EvaGreen was used to analyze methylation differences at the SNRPN locus in anonymized DNA samples from individuals with PWS (n = 39) or AS (n = 31) and from healthy control individuals (n = 95). Results from the MS-HRM assay were compared to those obtained by use of a methylation-specific PCR (MSP) protocol that is used commonly in diagnostic practice. RESULTS: With the MS-HRM assay 97.6% of samples were unambiguously assigned to the 3 diagnostic categories (AS, PWS, normal) by use of automated calling with an 80% confidence percentage threshold, and the failure rate was 0.6%. One PWS sample showed a discordant result for the MS-HRM assay compared to MSP data. CONCLUSIONS: MS-HRM is a simple, rapid, and robust method for screening methylation differences at the SNRPN locus and could be used as a diagnostic screen for PWS and AS
This record has no associated files available for download.
More information
Published date: 1 November 2007
Keywords:
chromosomes, syndrome, methods, deficiency, polymerase chain reaction, genotype, research support, genetics, uniparental disomy, diagnosis, angelman syndrome, ribonucleoproteins, autoantigens, human, pair 15, metabolism, analysis, transition temperature, prader-willi syndrome, antigens, dna methylation, laboratories, dna, research, small nuclear, humans
Identifiers
Local EPrints ID: 60421
URI: http://eprints.soton.ac.uk/id/eprint/60421
PURE UUID: dfd8c9b8-040a-475f-aae0-634bbd273882
Catalogue record
Date deposited: 08 Sep 2008
Last modified: 16 Mar 2024 03:23
Export record
Altmetrics
Contributors
Author:
Helen E. White
Author:
Victoria J. Hall
Download statistics
Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.
View more statistics
