Relevance of Polymerase Chain Reaction technique in immunogenetics and histocompatibility

Abstract

During the last decade of the previous century, among many of developing sciences, molecular biology had the most important position. The greatest influence on the development of scientific knowledge had been brought by discovery of PCR technique, that showed numerous advantages in comparison to the existing techniques. The capability of PCR technique to amplify specific DNA sequences, in a simple, automatic procedure, accelerated the development of many fields in molecular biology researches such as cloning, identification of new genes and pathogenes, determination and quantification of nucleotide sequences, thus opening fields of experimental investigations and its clinical application. PCR technique is widely used in various investigations from anthropological researches to those in molecular biology, from basic genetic investigations to clinical diagnostics. In the field of immunogenetics and histocompatibility, there are several HLA genotyping techniques: PCR-SSOP, detection by sequence specific oligonucleotide probes. PCR-SSP, detection by sequence specific primers, RSCA, detection by Reference-Strand Conformation Analysis, SBT-Sequence Based Typing. The development of these techniques has enabled the testing of the HLA alleles as well as detection of shared nucleotide sequences between alleles on the same or/and different loci and the existence of some locus-specific nucleotide sequences in coding (exon) and noncoding (intron) regions. The application of PCR technique in immunogenetics made it possible to define genetic polymorphism in HLA system, which contributed to a significant progress in the field of tissue and organ transplantation, population and genetic studies as well as in studies of disease association and HLA alleles.