| 摘要 |
FIELD: medicine.SUBSTANCE: group of inventions refers to medicine, namely to immunology, and can be used for diagnosing the condition of patient's immune system based on estimating number of T and B naive cells expressing TREC and KREC. For this purpose, DNA is recovered from analysed blood sample. Then, simultaneous amplification of sections of DNA molecules TREC, KREC and normalization locus IL17RA constantly present in single instance in each cycle of amplification, carried out by means of polymerase chain reaction (PCR) performed in common vessel at the same conditions for all amplified molecules and with participation of oligonucleotide fluorescence marked probes, carriers on the 5′-end fluorofore and 3′-end nefluorescent extinguisher, complementary to central part of amplified fragments. Obtained results are recorded by hybridization-fluorescent detection and determining in real time absolute number of molecules TREC, KREC and normalization locus IL17RA in analysed sample of DNA by means of calibration curves plotted in amplification of standard DNA samples taken in at least 4 serially decreasing ten times dilutions of known concentrations. Results of assessment of their quantity is diagnosing state of patient's immune system. Group of inventions also refers to set of primers, probes and standard samples for estimating number of DNA molecules TREC, KREC, IL17RA and number of genome DNA equivalents.EFFECT: using this group of inventions enables to determine the number of TREC and KREC of naive T and B cells in blood samples using set of declared primers.5 cl, 3 tbl, 4 ex, 4 tbl |
