High-resolution DNA melting analysis in clinical research and diagnostics

Abstract

Among nucleic acid analytical methods, high-resolution melting analysis is gaining more and more attention. High-resolution melting provides simple, homogeneous solutions for variant scanning and genotyping, addressing the needs of today's overburdened laboratories with rapid turnaround times and minimal cost. The flexibility of the technique has allowed it to be adopted by a wide range of disciplines for a variety of applications. In this review we examine the broad use of high-resolution melting analysis, including gene scanning, genotyping (including small amplicon, unlabeled probe and snapback primers), sequence matching and methylation analysis. Four major application arenas are examined to demonstrate the methods and approaches commonly used in particular fields. The appropriate usage of high-resolution melting analysis is discussed in the context of known constraints, such as sample quality and quantity, with a particular focus placed on proper experimental design in order to produce successful results.

Mutation Analysis of SLC26A4 for Pendred Syndrome and Nonsyndromic Hearing Loss by High-Resolution Melting

“…(exon 10) and c.1615-2AϾG (intron 14); P2 was heterozygous for p.Thr485Met (c.1454CϾT) (exon 13); and P3 was heterozygous for p.Thr721Met (c.2162CϾT) (exon 19). No other variants were detected in these samples, indicating that the HRM results were completely consistent with the Sanger sequencing results obtained during routine clinical testing.…”

“…It has the advantage of detecting heterozygous and homozygous sequence changes by monitoring the changes in melting temperature and melting curve shape compared with a reference sample. [17][18][19] With the addition of an intercalating dye in the initial PCR, HRM signals can be collected immediately after PCR amplification, without any post-PCR modification. Therefore, this simple and fast closed-tube system is especially suitable for clinical diagnostic applications.…”

Role of Melt Curve Analysis in Interpretation of Nutrigenomics’ MicroRNA Expression Data

“…HRM has been used to identify variation in nucleic acid sequences, enabled by use of a more advanced software, and is therefore less expensive than probe-based genotyping methods, and allows for identification of variants quickly and accurately (75). This method has been widely used in molecular diagnosis and for detection of mutations (76)(77)(78).…”

Targeted mutation breeding of horticultural plants

“…Improvement of mismatch detection by HRM and TILLING is an active field of study, largely due to medical diagnostic applications (e.g. Li et al, 2008;Montgomery et al, 2010).…”