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Development of a new genotyping assay for detection of the BDNF Val66Met polymorphism using melting-curve analysis

Abstract:

Brain-derived neurotrophic factor (BDNF) plays critical role in the growth, differentation and survival of neurons in the CNS. Recent research has suggested that BDNF may be implicated in the etiology of mood disorders and schizophrenia, as well as in the therapeutic action of some drugs, such as antidepressants and antipsychotics. This study aimed to develop a simple, fast and accurate new methods for detecting the Va166Met polymorphism of the BDNF gene in schizophrenis patients using melting-curve analysis and a DNA-specific dye, SYBR® Green I. A group of 30 schizoprenia patients were analyzed to detect the BDNF Va166Met polymorphim (rs6265) using the new genotyping methods based on the analysis of fluorescence melting curves of PCR products that were labeled with SYBR Green I. The genotype results were confirmed for all 30 samples using the specific BDNF TaqMan® allele discrimination assay. This new methods allows the analysis of both alleles in the same reaction tube using SYBR Green I, with no need for additional steps. The addition of a GC clamp makes this methods universally applicable, since the melting temperature of one allele can be adjusted as necessary to give the distinctive separation of melting curve. Therefore, this new methods is simple, fast and accurate for determining the presence of the BDNF Va166Met polymorphism. It may also be useful for the analysis of other SNPs in pharmacogenetic studies. © 2009 Future Medicine Ltd.