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Method and kit for genotype detection of CYP3a5*3 sites

A genotype and kit technology, applied in the field of genotype detection and kits for CYP3A5*3 loci, can solve the problems of complex therapeutic index, narrow pharmacokinetic high variability, etc., and reduce the occurrence of adverse reactions Effect

Inactive Publication Date: 2018-06-19
GUANGZHOU HEKANG MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Its clinical use is complicated by the high variability among its pharmacokinetics and its narrow therapeutic index

Method used

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  • Method and kit for genotype detection of CYP3a5*3 sites
  • Method and kit for genotype detection of CYP3a5*3 sites
  • Method and kit for genotype detection of CYP3a5*3 sites

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The invention provides a kit, which includes a primer with the following sequence:

[0024]

[0025] Specific probes for the following sequences:

[0026]

[0027] Reporter probe for the following sequence: tatctcttcc ctgtttggac.

Embodiment 2

[0029] A method for genotype detection of CYP3A5*3 site, the method includes the steps of: performing PCR reaction in a reaction tube, the reaction system has a total volume of 10 μl, including 2x qiagen Hotstar MM 5ul, primer mix 1ul, DNA sample 2ul , Sterilized water 2ul.

[0030] The reaction was carried out on the ABI9700 PCR amplification instrument. The reaction conditions were 95°C, 15min, and 30 cycles of 94°C, 30 seconds, 60°C, 30 seconds, 72°C, 30 seconds; 72°C, 7 minutes, 4°C maintain.

[0031] Multiplex OLA reaction: Prepare 2xOLA master mix: 10x Taq Ligase buffer 2ul, Taq DNALigase (40,000U / ml) 0.25ul, wild-type probe mix (100nM each) 1ul, mutant probe mix (2.5uMeach) 2ul, deionized Water 4.75ul. Mix OLA master mix with the reaction product: 2xOLA master mix 10ul, amplified PCR product 5ul, sterilized deionized water 5ul. Pipette up and down to mix well, cover the reaction tube, and carry out the ligation reaction on the ABI9700 PCR amplification instrument. 96°...

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Abstract

The invention provides a method and a kit for genotype detection of CYP3a5*3 sites and relates to the field of molecular biology and medicine. Specific primers, probes and magnetic beads written intorelated report probes are designed for gene SNP sites related to risk of adverse reactions associated with dosage of tacrolimus and cyclosporine, and types of the SNP sites can be subjected to typingdetection. By means of the detection kit, the type of CYP3A5*3(G >A) in human whole blood samples can be detected sensitively and rapidly, reliable experimental evidence for application dosage adjustment of tacrolimus and cyclosporine can be provided for patients, adverse reactions are reduced, and clinical treatment is guided.

Description

technical field [0001] The present invention relates to the fields of molecular biology and medicine. More specifically, the present invention relates to a kit of SNP sites related to the risk of adverse reactions of tacrolimus and cyclosporine. The single nucleotide polymorphism site (SNP) genotype of cytochrome oxidase CYP3A5*3 gene was used to evaluate the adverse reactions and dose adjustment of tacrolimus and cyclosporine during application. Background technique [0002] Cytochrome P450 3A (CytochromeP450 3A, CYP3A) is the most important CYP450 enzyme in the body, accounting for 25% of the total CYP450 enzymes in the adult liver, and is also an important CYP enzyme in the intestine, mainly composed of CYP3A4, CYP3A5, CYP3A7 and CYP3A43, of which CYP3A4 and CYP3A5 are especially important. CYP3A4 and CYP3A5 catalyze the oxidation reaction of more than 60% of commonly used clinical drugs. There are obvious individual differences in CYP3A gene expression. In adults, the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/106C12Q2600/156C12Q2600/158
Inventor 杜予和
Owner GUANGZHOU HEKANG MEDICAL TECH CO LTD
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