SNPs Genotyping by High Resolution Melt Analysis
Results of Human Genome Project have brought researchers closer to understand the relationship between individual genetic constitution and drug response differences among populations, as well as diseases susceptibility. The ability to detect highly predictive genotype-phenotype association may be useful for various applications, from phisicians to identify the best therapy, to epidemiologists for association studies till researchers involved in new drug development. Only in 2010, pharmacogenetics/pharmacogenomics and SNPs topics are covered respectively in over than 5000 and 700 publications in the NCBI database. Pharmacogenetics is evolved from a niche discipline to a major driving force in clinical pharmacology and currently it is one of the most actively pursued disciplines in applies biomedical research. [Jurgen Brockmoller & Mladen V. Tzvetkov, 2008] Pharmacogenetics has changed the preclinical and clinical practices in drug research and today, large clinical trials without a pharmacogenetic add-on, appear to have become the minority [Jurgen Brockmoller & Mladen V. Tzvetkov, 2008]. Moreover, the advances in high-throughput genotyping technologies are poised to provide a flood of information that will affect both pharmacogenetic discovery and pharmacogenetic application in clinical practice. Although direct DNA sequencing is considered as a "gold standard" for genotyping of SNPs and mutations, it still remains relatively expensive, laborious and time consuming [Patricia A. et al 2009] . Different methods have been developed to simplify the detection of novel mutations, from RFLP to SSCP ( single strand conformation polymorphism) and DGGE (denaturing gradient gel electrophoresis), DHPLC (denaturing high performance liquid chromatography) DNA microarray, and finally several PCR based methods (allele/specific amplification, single-base extension etc).
High Resolution Melting (HRM) is a simple, rapid and low-cost mutation scanning method. Its advantage is that PCR amplification and melting curve analysis are performed within the same tube or plate, without any post-PCR processing; these features are particularly important for a routine diagnostic setting.
HRM is today one of the most cost-efficient method for SNPs genotyping
Diatheva is committed to developing kits for SNPs detection in the field of pharmacogenetics and predictive medicine.
In order to simplify and speed up researcher and technicians work, our kits are made to economize as much as possible both time and manual labour and to minimize the needs of results interpretation. Considering the possibility that one SNP can be investigated in associatioon with several others, it is essential to provide a flexible platform that allows simultaneous detection of several SNPs. In order to reach this aim our kits are developed sharing the same operating conditions, with optimized and ready-to-use Master Mix and specific thermal profiles, enabling clustering of several SNPs and their simultaneous detection.
All kits are developed and validated using Rotor Gene 6000 (Corbett Research) Rotor Gene Q (Qiagen).
Diatheva provides a production service on demand for customized HRM kits CE marked and protocols for SNPs detection by HRM.