FIELD: medicine.
SUBSTANCE: method of whole blood HLA-typing refers to biotechnology. The method involves DNA recovery from blood with using a set of reagents of various ELB A, ELB B buffer solutions and centrifugation at various accelerations; a blood sample 0.7 ml is washed by removing erythrocytes and residues; leukocytes are lysed by means of the TBS solution; DNA is precipitated on a membrane from the same set and mechanically processed by centrifugation to produce high-purity DNA. The DNA concentration is checked on the DU 800 spectrophotometer with using bidistilled water by comparing a purity value of the latter with purity of washed DNA. Blood genetic typing is performed by molecular-genetic method of polymerase chain reaction (PCR) by placing a plate with frozen-dried primers from the set Protrans HLA-A*, B*, DRBI* on the PROTRANS workstation with using mixed D, Y and Taq polymerase buffers and DNA samples, with performing an amplification procedure in a thermal cycle apparatus by the preset instruction with the specified parameters. The PCR results are visualised by an electrophoresis method with agarose gel painted by ethidium bromide placed in an electrophoresis cell. Under current, DNA molecule pass through the gel. The presence of the product in the plate well with specific primers shows a HLA-genotype.
EFFECT: invention provides higher reliability and quality of the typing results.
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