Nöropsikiyatrik ilaç kullanımında yeni nesil dizileme yöntemi ile tanı paneli geliştirilmesi / The development of pharmacogenetics diagnostic panels with next generation sequencing methods in neuropsychatric drug use

Abstract

As in the whole world, health expenditures have increased as a result of the use of high-tech technological developments in our country, and at the same time, we have been able to make some savings in spending with developing technology. Pharmacogenetic tests will be the most effective assistant to this saving. The share of medicines in health expenditures in Turkey 46%, while in Britain and America, this ratio is only 12%. Pharmacogenetics is a scientific discipline that examines changes in the responses people give to drugs and the reasons for variations in genetic constitution. After the completion of the Human Genome Project in 2003 and is spreading rapidly in the world. By comparing the genetic structure of the individual with the drugs that can be used in the treatment of the disease, it is ensured that the side effect of the most effective drug can be given in the least possible dose. Nowadays pharmacogenetic tests can be performed on individuals before treatment of many diseases such as epilepsy, diabetes, heart diseases, stomach diseases, cancer and depression. In the future of pharmacogenetics, three areas appear to be particularly prominent. These areas are psychiatry, cardiology and oncology. In 2000, depression was ranked fourth among the causes of the global illness burden, which was assessed as '' Disability-adjusted life year (DALY) ''. According to the World Health Organization (WHO) reports, by the year 2020, depression will take the second place in the " Disability-adjusted life year" evaluations for all age groups and gender groups. While 35-45% of the drugs used by our psychiatric patients respond to treatment, 30-50% of them produce severe side effects that either do not respond to treatment or interfere with the flow of daily life. Nearly all drugs used in the field of neuropsychiatry are metabolized by the enzymes CYP2D6, CYP2C9, CYP2C19, CYP1A2 and CYP3A4, which are produced in the liver and have high polymorphism Although not widely used, polymorphisms in these genes can be detected by DNA analysis methods and appropriate drugs and doses can be determined before treatment begins. These analyzes are made using different methods and no standardization has been achieved yet. PCR-RFLP, Sanger DNA sequence analysis, Real-time PCR, microarray, etc. The purpose of the project is to create a molecular diagnostic kit that provides analysis of all of these genes, which are known to be routinely applied, based on polymorphisms and mechanisms of action according to the literature. As a method, it was planned to replicate 52 polymorphic spots with 22 primers which are most frequently used in pharmacogenetic analysis for neuropsychiatric genes in genes encoding the target enzymes. Suitable primers were designed and optimized to allow analysis of points. Amplification of the genes was achieved by PCR using the optimized primers. Later, sequence analyzes were performed with the next generation of scaling technology (Illumina platform). After the quality analysis of the data, the polymorphisms they carried out were determined after the scaling process. The study was not a community survey through a polymorphism, but a polymorphism analysis panel / kit was developed to perform analysis of many SNP at once. Genomic DNA of 20 different individuals randomly selected was used for the method development. The kit designed, more economical and next generation technology for clinical use has been developed.