SET OF REAGENTS FOR THE DETERMINATION OF RIBONUCLEIC ACIDS IN THE DIAGNOSIS OF LYMPHOPROLIFERATIVE DISEASES Russian patent published in 2023 - IPC C12Q1/68 

FIELD: medicine.

SUBSTANCE: invention relates to clinical laboratory diagnostics of lymphocytic lymphoma/chronic lymphocytic leukemia (CLL) using molecular genetic methods for simultaneously determining the level of expression of ribonucleic acids of different classes: microRNA and matrix RNA in a multiplex polymerase chain reaction in the "real time mode". The invention is a kit containing reagents for RNA isolation, reverse transcription reagents, including specific loop primers for microRNA, reagents for PCR, including optimally selected reaction mixtures for determining the expression of ABL1 mRNA and ROR1 mRNA, as well as miR-39 microRNA, miR-155 and miR-15a, Thermus aquaticus DNA polymerase, a positive control, a negative control, and a control marker for miRNA extraction efficiency. A distinctive feature of the kit is the use of original combinations of specific primers and fluorescently labeled DNA probes in three reaction mixtures, as well as the possibility of simultaneous determination of mRNA and microRNA. The first mixture includes the following primers: ABL-F, ABL-R, ROR-F, ROR-R and TaqMan probes: ABL-P, ROR-P with dyes. The second mixture contains the following primers: miR155-F, miR155-R, miR39-R, miR39-F and TaqMan probes: miR155-P, miR39-P with dyes. The third mixture contains the following primers: miR15a-F, miR15a-R. The invention allows simultaneous determination of the level of mRNA and microRNA in one analytical series using three test tubes with reaction mixtures. The technical result achieved by the claimed invention is a set of reagents for multiplex polymerase chain reaction, which allows to perform molecular genetic diagnosis of CLL using standard equipment of clinical diagnostic laboratories.

EFFECT: using the capabilities of the multiplex reaction mode leads to savings in the volume of the biological sample used, consumables and reduces the time of the analysis.

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