IMMUNODIAGNOSTIC METHOD AND SET FOR THE ANALYSIS OF DNA MOLECULES OF TREC AND KREC AND OF THE NUMBER OF DNA GENOME EQUIVALENTS

摘要

The group of inventions relates to the field of medical and clinical diagnostics and can be used for the quantitative analysis of TREC and KREC molecules, which are markers of naive T and B cells respectively, in order to determine the condition of a patient's immune system. The present method involves isolating DNA from a sample of blood under analysis and simultaneously amplifying regions of DNA molecules of TREC and KREC and a reference locus which is constantly present in single quantities in each cycle of a polymerase chain reaction (PCR) amplification procedure. The results obtained are recorded using hybridization-fluorescence detection, the absolute number of TREC, KREC and reference locus molecules in the DNA sample under analysis is determined in real time with the aid of calibration curves, and the results of the quantitative analysis of said molecules are used to diagnose the condition of the patient's immune system. Also provided is a set for the real-time quantitative analysis of DNA molecules of TREC and KREC and of the number of DNA genome equivalents using PCR, and also a set of reference plasmid DNA samples with a precisely determined absolute concentration. The invention makes it possible to reduce errors in determining the concentration of TREC and KREC molecules, particularly with regard to low values, and also reduces the number of reactions necessary for analysis by virtue of multiplexing.