Significance of Methods for Purification of Oligodeoxyribonucleotide Probes for the Efficiency of Gene Diagnosis by Real-Time PCR

Analysis of the use of real-time PCR with fluorescent registration of results for gene diag-nosis of infectious diseases showed that the sensitivity and reliability of quantitative eva-luation of DNA targets directly depended on the method of purification of oligonu-cleotide probes. Chromatographic behavior of synthetic probes carrying various fluoro-phores and fluorescence quenchers was analyzed. Approaches to optimization of purification methods are proposed enabling elimination of previously undetectable admix-tures. The importance of these studies is explained by the need in extending the armory of methods for the development and production of diagnosticums for detection of infectious and hereditary diseases, identification of genetically modified organisms, and for a wide spectrum of research in molecular biology and medicine. Key Words: oligodeoxyribonucleotide probes; polymerase chain reaction in the real-time mode; gene diagnosis; high performance liquid chromatography; mass spectrometr