Method and kit for detecting SNP locus genotypes having adverse reactions to CTX drugs

A technology of adverse reactions and genotypes, applied in biochemical equipment and methods, microbiological determination/inspection, etc., can solve the problems of high risk of adverse reactions, and achieve the effect of guiding clinical treatment selection, rapid detection, and reliable experimental evidence

Inactive Publication Date: 2018-05-08
GUANGZHOU HEKANG MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The MTHFRC677T polymorphism is associated with increased toxicity, and patients carrying the T allele at the C677T locus are at high risk of adverse reactions

Method used

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  • Method and kit for detecting SNP locus genotypes having adverse reactions to CTX drugs
  • Method and kit for detecting SNP locus genotypes having adverse reactions to CTX drugs
  • Method and kit for detecting SNP locus genotypes having adverse reactions to CTX drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] The invention provides a kit for detecting the genotype of the SNP locus of an adverse drug reaction to CTX. The kit includes primers with the following sequences:

[0026]

[0027]

[0028] Specific probe sequences for the following sequences:

[0029]

[0030] Specific reporter gene for the following sequence:

[0031]

Embodiment 2

[0033] The 3 SNP sites were subjected to PCR reaction in a PCR reaction tube, and the reaction system had a total volume of 10 μl, including 2xqiagen Hotstar MM 5ul, primer mix 1ul, DNA sample 2ul, and sterilized water 2ul.

[0034] 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.

[0035] 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 ligation reaction on ABI9700 PCR amplification instrument: 96°C for 2min, 30 cycles of 94°C for 15s, 37°C for 1min; maint...

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Abstract

The invention discloses a method and kit for detecting SNP locus genotypes having adverse reactions to CTX drugs. The invention relates to the fields of molecular biology and medicine; according to the kit provided by the invention, aiming at SNP loci with cyclophosphamide adverse reaction risks and in human genomic DNA, namely a GSTP1 gene rs1695, specific primers and probes are designed, a hybridization reaction with MagPlex-TAG magnetic beads with corresponding report genes is performed, and detection is performed on a luminex 200 instrument through chromogenic reaction; a base type of eachSNP locus is judged through reading and interpretation of signal values. The kit has the beneficial effects that the specific primers, probes and report gene probe sequences are designed by using theSNP loci related to the adverse reaction risks of cyclophosphamide used by patients with systemic lupus erythematosus and other diseases, and the types of the SNP loci can be detected in a typing manner. The detection kit provided by the invention can detect the types of the SNP sensitively and quickly, and can provide reliable experimental evidence for predicting medication adverse reaction risks of the patients with systemic lupus erythematosus, and the clinical treatment selection 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 set of kits for cyclophosphamide drug adverse reaction risk SNP sites, through simultaneous detection of GSTP1 gene and MTHFR gene which are closely related to the risk of cyclophosphamide toxicity and side effects The single nucleotide polymorphism site (SNP) genotype was used to assess the risk of adverse effects of cyclophosphamide in patients with systemic lupus erythematosus and other diseases. Background technique [0002] SLE is a common autoimmune disease. Cyclophosphamide plays an important role in the treatment of SLE and is an effective drug for severe SLE. Glucocorticoid combined with high-dose cyclophosphamide has become the standard treatment for systemic lupus erythematosus or lupus nephritis. Cyclophosphamide can improve the condition of SLE and is a strong cellular immunosuppressant, but CTX is more contr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/6827
CPCC12Q1/6827C12Q1/6883C12Q2600/106C12Q2600/156C12Q2600/16C12Q2600/166C12Q2531/113C12Q2537/143C12Q2563/143C12Q2563/149
Inventor 杜予和
Owner GUANGZHOU HEKANG MEDICAL TECH CO LTD
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