High resolution melting analysis facilitates mutation screening of ETFDH gene: Applications in riboflavin-responsive multiple acyl-CoA dehydrogenase deficiency

2010 ◽  
Vol 411 (9-10) ◽  
pp. 690-699 ◽  
Author(s):  
Tze-Kiong Er ◽  
Wen-Chen Liang ◽  
Jan-Gowth Chang ◽  
Yuh-Jyh Jong
Keyword(s):  
High Resolution ◽  
Dehydrogenase Deficiency ◽  
High Resolution Melting ◽  
Mutation Screening ◽  
High Resolution Melting Analysis ◽  
Melting Analysis

scholarly journals NF2 Mutation Screening by Denaturing High-Performance Liquid Chromatography and High-Resolution Melting Analysis

2008 ◽  
Vol 12 (2) ◽  
pp. 311-318 ◽  
Author(s):  
Roberta Sestini ◽  
Aldesia Provenzano ◽  
Costanza Bacci ◽  
Claudio Orlando ◽  
Maurizio Genuardi ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Resolution ◽  
High Performance ◽  
High Resolution Melting ◽  
Mutation Screening ◽  
High Resolution Melting Analysis ◽  
Melting Analysis

Impact of fixation artifacts and threshold selection on high resolution melting analysis for KRAS mutation screening

2017 ◽  
Vol 35 ◽  
pp. 34-43 ◽  
Author(s):  
Wendy Pérez-Báez ◽  
Ethel A. García-Latorre ◽  
Héctor Aquiles Maldonado-Martínez ◽  
Iris Coronado-Martínez ◽  
Leonardo Flores-García ◽  
...  
Keyword(s):  
High Resolution ◽  
Kras Mutation ◽  
High Resolution Melting ◽  
Mutation Screening ◽  
High Resolution Melting Analysis ◽  
Threshold Selection ◽  
Melting Analysis

scholarly journals Mutation Scanning the GJB1 Gene with High-Resolution Melting Analysis: Implications for Mutation Scanning of Genes for Charcot-Marie-Tooth Disease

2007 ◽  
Vol 53 (2) ◽  
pp. 349-352 ◽  
Author(s):  
Marina L Kennerson ◽  
Trent Warburton ◽  
Eva Nelis ◽  
Megan Brewer ◽  
Patsie Polly ◽  
...  
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Mutation Screening ◽  
Gene Mutations ◽  
High Resolution Melting Analysis ◽  
Charcot Marie Tooth ◽  
Charcot Marie Tooth Disease ◽  
Mutation Scanning ◽  
Junction Protein ◽  
Melting Analysis

Abstract Background: X-linked Charcot-Marie-Tooth type 1 disease has been associated with 280 mutations in the GJB1 [gap junction protein, beta 1, 32kDa (connexin 32, Charcot-Marie-Tooth neuropathy, X-linked)] gene. High-resolution melting analysis with an automated instrument can be used to scan DNA for alterations, but its use in X-linked disorders has not been described. Methods: A 96-well LightScanner for high resolution melting analysis was used to scan amplicons of the GJB1 gene. All mutations reported in this study had been confirmed previously by sequence analysis. DNA samples were amplified with the double-stranded DNA-binding dye LC Green Plus. Melting curves were analyzed as fluorescence difference plots. The shift and curve shapes of melting profiles were used to distinguish controls from patient samples. Results: The method detected each of the 23 mutations used in this study. Eighteen known mutations provided validation of the high-resolution melting method and a further 5 mutations were identified in a blind study. Altered fluorescence difference curves for all the mutations were easily distinguished from the wild-type melting profile. Conclusion: High-resolution melting analysis is a simple, sensitive, and cost-efficient alternative method to scan for gene mutations in the GJB1 gene. The technology has the potential to reduce sequencing burden and would be suitable for mutation screening of exons of large multiexon genes that have been discovered to be associated with Charcot Marie Tooth neuropathy.

scholarly journals Detection of Factor VIII Gene Mutations by High-Resolution Melting Analysis

2007 ◽  
Vol 53 (12) ◽  
pp. 2211-2214 ◽  
Author(s):  
Andrew D Laurie ◽  
Mark P Smith ◽  
Peter M George
Keyword(s):  
High Resolution ◽  
Dna Sequencing ◽  
Factor Viii ◽  
High Resolution Melting ◽  
Hemophilia A ◽  
Mutation Screening ◽  
Gene Mutations ◽  
High Resolution Melting Analysis ◽  
Melting Curves ◽  
Melting Analysis

Abstract Background: Single base-pair substitution mutations in the gene for coagulation factor VIII, procoagulant component (hemophilia A) (F8) account for approximately 50% of severe cases of hemophilia A (HA), and almost all moderate or mild cases. Because F8 is a large gene, mutation screening using denaturing HPLC or DNA sequencing is time-consuming and expensive. Methods: We evaluated high-resolution melting analysis as an option for screening for F8 gene mutations. The melting curves of amplicons heterozygous for known F8 gene mutations were compared with melting curves of the corresponding normal amplicons to assess whether melting analysis could detect these variants. We examined 2 platforms, the Roche LightCycler 480 (LC480) and the Idaho Technology LightScanner. Results: On both instruments, 18 (90%) of the 20 F8 gene variants we examined were resolved by melting analysis. For the other 2 mutations, the melting curves of the heterozygous amplicons were similar to the corresponding normal amplicons, suggesting these variants may not be detected by this approach in a mutation-scanning screen. Conclusion: High-resolution melting analysis is an appealing technology for F8 gene screening. It is rapid and quickly identifies mutations in the majority of HA patients; samples in which no mutation is detected require further testing by DNA sequencing. The LC480 and LightScanner platforms performed similarly.

scholarly journals Parental mosaicism in Marfan and Ehlers–Danlos syndromes and related disorders

2021 ◽  
Author(s):  
Bertrand Chesneau ◽  
Aurélie Plancke ◽  
Guillaume Rolland ◽  
Nicolas Chassaing ◽  
Christine Coubes ◽  
...  
Keyword(s):  
High Resolution ◽  
Marfan Syndrome ◽  
High Resolution Melting ◽  
De Novo ◽  
Direct Sequencing ◽  
Causal Variant ◽  
Connective Tissue Disorder ◽  
High Resolution Melting Analysis ◽  
Aortic Diseases ◽  
Melting Analysis

AbstractMarfan syndrome (MFS) is a heritable connective tissue disorder (HCTD) caused by pathogenic variants in FBN1 that frequently occur de novo. Although individuals with somatogonadal mosaicisms have been reported with respect to MFS and other HCTD, the overall frequency of parental mosaicism in this pathology is unknown. In an attempt to estimate this frequency, we reviewed all the 333 patients with a disease-causing variant in FBN1. We then used direct sequencing, combined with High Resolution Melting Analysis, to detect mosaicism in their parents, complemented by NGS when a mosaicism was objectivized. We found that (1) the number of apparently de novo events is much higher than the classically admitted number (around 50% of patients and not 25% as expected for FBN1) and (2) around 5% of the FBN1 disease-causing variants were not actually de novo as anticipated, but inherited in a context of somatogonadal mosaicisms revealed in parents from three families. High Resolution Melting Analysis and NGS were more efficient at detecting and evaluating the level of mosaicism compared to direct Sanger sequencing. We also investigated individuals with a causal variant in another gene identified through our “aortic diseases genes” NGS panel and report, for the first time, on an individual with a somatogonadal mosaicism in COL5A1. Our study shows that parental mosaicism is not that rare in Marfan syndrome and should be investigated with appropriate methods given its implications in patient’s management.

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