scholarly journals Diagnosis of ABCB11 gene mutations in children with intrahepatic cholestasis using high resolution melting analysis and direct sequencing

2014 ◽  
Vol 10 (3) ◽  
pp. 1264-1274 ◽  
Author(s):  
GUORUI HU ◽  
PING HE ◽  
ZHIFENG LIU ◽  
QIAN CHEN ◽  
BIXIA ZHENG ◽  
...  
Keyword(s):  
High Resolution ◽  
Intrahepatic Cholestasis ◽  
High Resolution Melting ◽  
Direct Sequencing ◽  
Gene Mutations ◽  
High Resolution Melting Analysis ◽  
Melting Analysis

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.

Identification of fibrillin-1 gene mutations in Marfan syndrome by high-resolution melting analysis

2009 ◽  
Vol 389 (2) ◽  
pp. 102-106 ◽  
Author(s):  
Chia-Cheng Hung ◽  
Shin-Yu Lin ◽  
Chien-Nan Lee ◽  
Hui-Yu Cheng ◽  
Chiou-Ya Lin ◽  
...  
Keyword(s):  
High Resolution ◽  
Marfan Syndrome ◽  
High Resolution Melting ◽  
Gene Mutations ◽  
High Resolution Melting Analysis ◽  
Fibrillin 1 ◽  
Melting Analysis

Detection of SF3B3 Gene Mutations in Oral Cancer by High Resolution Melting Analysis

2014 ◽  
Vol 60 (12/2014) ◽  
Author(s):  
Chun-Chi Chang ◽  
Ya-Sian Chang ◽  
Wen-Ling Chan ◽  
Kun-Tu Yeh ◽  
Ren-Jeng Wei ◽  
...  
Keyword(s):  
High Resolution ◽  
Oral Cancer ◽  
High Resolution Melting ◽  
Gene Mutations ◽  
High Resolution Melting Analysis ◽  
Melting Analysis

EGFR mutations detected by high-resolution melting analysis (HRMA) as a predictor of response and survival in non-small cell lung cancer (NSCLC) patients treated with gefitinib

2006 ◽  
Vol 24 (18_suppl) ◽  
pp. 7075-7075
Author(s):  
T. Takano ◽  
Y. Ohe ◽  
K. Furuta ◽  
K. Tsuta ◽  
K. Nomoto ◽  
...  
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Response Rate ◽  
Direct Sequencing ◽  
False Negative ◽  
High Resolution Melting Analysis ◽  
Egfr Mutations ◽  
Wild Type ◽  
Minor Response ◽  
Melting Analysis

7075 Background: Recent studies have shown that EGFR mutations, mainly deletions in exon 19 (DEL) and L858R, are associated with gefitinib sensitivity in patients (pts) with NSCLC. We established a new easy method, using high-resolution melting analysis (HRMA), for detecting DEL and L858R mutations even from small biopsy or cytology samples, and evaluated the significance of EGFR mutations in NSCLC on a larger scale. Methods: Among 364 advanced or recurrent NSCLC pts treated with gefitinib between Jul 2002 and Dec 2004, HRMA was performed in 207 pts from whom specimens were available. DNA extracted from the archival tissue or cytology samples not subjected to microdissection was analyzed to detect DEL and L858R using HR-1 (Idaho Technology), an HRMA device. To validate this method, the results were compared with direct sequencing data obtained from microdissected tumor cells from surgical specimens in 66 pts. Results: Tissue/cytology/both samples were analyzed in 91/77/39 pts. EGFR mutations were detected in 85 (41%; DEL/L858R: 49/36) of the 207 pts. In the comparison with direct sequencing, consistent results were obtained from all of the 66 tissue samples, while false negative results were obtained in 2 of the 28 cytology samples. EGFR mutations were seen more frequently in women (54% vs. 31%; P = .001), never-smokers (53% vs. 32%; P = .002), and pts with adenocarcinoma (44% vs. 11%; P = .007). CR/PR/SD/PD was observed in 2/64/11/8 pts with EGFR mutations and in 0/10/23/89 pts with wild-type EGFR. The response rate (78% vs. 8%), time to progression (median, 9.1 vs. 1.6 months) and overall survival (median, 19.9 vs. 9.1 months) were all significantly superior in pts with EGFR mutations (P < .0001). Minor response and/or long SD (>6 months) was observed more frequently in SD pts with EGFR mutations than in those with wild-type EGFR (91% vs. 26%; P < .001). Among the pts with EGFR mutations, the response rate was significantly higher in the pts with DEL than in those with L858R (86% vs. 67%; P = .037). Conclusions: HRMA is a practical and precise method to detect DEL and L858R mutations. EGFR mutations strongly predict a better response and longer survival in NSCLC pts treated with gefitinib. No significant financial relationships to disclose.

Rapid identification of HBB gene mutations by high-resolution melting analysis

2009 ◽  
Vol 42 (16-17) ◽  
pp. 1667-1676 ◽  
Author(s):  
Hung-Chang Shih ◽  
Tze-Kiong Er ◽  
Tien-Jye Chang ◽  
Ya-Sian Chang ◽  
Ta-Chih Liu ◽  
...  
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Gene Mutations ◽  
Rapid Identification ◽  
High Resolution Melting Analysis ◽  
Melting Analysis

High resolution melting analysis for the detection of SLC25A13 gene mutations in Taiwan

2011 ◽  
Vol 412 (5-6) ◽  
pp. 460-465 ◽  
Author(s):  
Jing-Ting Lin ◽  
Kwang-Jen Hsiao ◽  
Chiung-Yu Chen ◽  
Cheng-Chung Wu ◽  
Shio-Jean Lin ◽  
...  
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Gene Mutations ◽  
High Resolution Melting Analysis ◽  
Melting Analysis

scholarly journals Rapid and Sensitive Detection of BRCA1/2 Mutations in a Diagnostic Setting: Comparison of Two High-Resolution Melting Platforms

2008 ◽  
Vol 54 (6) ◽  
pp. 982-989 ◽  
Author(s):  
Kim De Leeneer ◽  
Ilse Coene ◽  
Bruce Poppe ◽  
Anne De Paepe ◽  
Kathleen Claes
Keyword(s):  
High Resolution ◽  
High Resolution Melting ◽  
Direct Sequencing ◽  
Turnaround Time ◽  
High Resolution Melting Analysis ◽  
Coding Region ◽  
Brca1 And Brca2 ◽  
Diagnostic Laboratory ◽  
Sensitivity Setting ◽  
Melting Analysis

Abstract Background: High-resolution melting is an emerging technique for detection of nucleic acid sequence variations. Developments in instrumentation and saturating intercalating dyes have made accurate high-resolution melting analysis possible and created opportunities to use this technology in diagnostic settings. We evaluated 2 high-resolution melting instruments for screening BRCA1 and BRCA2 mutations. Methods: To cover the complete coding region and splice sites, we designed 112 PCR amplicons (136–435 bp), amplifiable with a single PCR program. LCGreen® Plus was used as the intercalating dye. High-resolution melting analysis was performed on the 96-well Lightscanner™ (Idaho Technology Inc.) and the 96-well LightCycler® 480 (Roche) instruments. We evaluated sensitivity by analyzing 212 positive controls scattered over almost all amplicons and specificity by blind screening of 22 patients for BRCA1 and BRCA2. In total, we scanned 3521 fragments. Results: All 212 known heterozygous sequence variants were detected on the Lightscanner by analysis on normal sensitivity setting. On the LightCycler 480, the standard instrument sensitivity setting of 0.3 had to be increased to 0.7 to detect all variants, decreasing the specificity to 95.9% (vs 98.7% for the Lightscanner). Conclusions: Previously, we screened BRCA1/2 by direct sequencing of the large exon 11 and denaturing gel gradient electrophoresis (DGGE) for all other coding exons. Since the introduction of high-resolution melting, our turnaround time has been one third of that with direct sequencing and DGGE, as post-PCR handling is no longer required and the software allows fast analyses. High-resolution melting is a rapid, cost-efficient, sensitive method simple enough to be readily implemented in a diagnostic laboratory.

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