Method and kit for detecting SNP sites related to tacrolimus and cyclosporin A individalized medication

Abstract

The invention relates to a method and kit for detecting SNP sites related to tacrolimus and cyclosporin A individalized medication. The detection method comprises the following steps: (1) designing specific primers aiming at CYP3A4, CYP3A5 and MDR1 genes; (2) performing specific polymerase chain reaction (PCR) amplification so as to obtain target fragments containing CYP3A4, CYP3A5 and MDR1 genes;(3) performing restrictive enzyme disgestion on PCR product fragments; (4) performing single base extension reaction; (5) performing desalting purification treatment; and (6) then detecting and analyzing gene sequences of target genes CYP3A4, CYP3A5 and MDR1 genes. The method for detecting SNP sites related to tacrolimus and cyclosporin A individalized medication is high in detection accuracy, high in experiment repeatability, great in flux and low in cost. The invention also provides a kit for detecting SNP sites related to tacrolimus and cyclosporin A individalized medication. The kit comprises a specific primer pair for amplifying CYP3A4, CYP3A5 and MDR1 genes. The kit for detecting SNP sites related to tacrolimus and cyclosporin A individalized medication has the advantages of simplified experimental procedure, short manual operation time, low difficulty and high experiment automation degree.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a detection method and a kit for SNP sites related to individualized medication of tacrolimus and cyclosporine A. Background technique [0002] Tacrolimus is a fermentation product isolated from Streptomyces tsukubaensis, and its chemical structure belongs to 23-membered macrolide antibiotics. Tacrolimus is a powerful new type of immunosuppressant, mainly by inhibiting the release of interleukin-2 (L-2), and comprehensively inhibits the function of T lymphocytes. In recent years, it has been the first-line drug for liver and kidney transplantation. Cyclosporin is a cyclic polypeptide containing only 11 amino acids extracted from mold fermentation products discovered by Borel in Switzerland in the late 1970s, which can effectively and specifically inhibit lymphocyte response and proliferation. In addition to being used in organ transplantation, cyclosporin A can also...

AI Technical Summary

Problems solved by technology

The polymorphism of the MDR1 gene leads to differences in gene expression among individuals, resulting in differences in the ability of P-gp to transport drugs from intracellular to extracellular, resulting in individual differences in multidrug resistance, thus affecting the bioavailability of drugs. also different

In the prior art, direct sequencing is used to detect CYP3A4, CYP3A5, and MDR1 gene polymorphisms, which is costly, complicated, and time-consuming; the hybridization chip detection method has the problems of high false positive rate, accurate annealing temperature, and low specificity; Taqman method probe preparation and supporting equipment are expensive and the accuracy is not high

The limitations of the above methods limit the practical application of CYP3A4, CYP3A5, MDR1 gene polymorphism detection

Images