Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Gene detection kit for detecting individual drug use of tacrolimus drug

A gene detection and tacrolimus technology, which is applied in the field of gene detection kits for the detection of individualized drug use of tacrolimus drugs, to achieve the effects of reducing drug side effects, realizing individualized treatment, and improving drug efficacy

Inactive Publication Date: 2019-10-22
上海佰臻生物科技有限公司
View PDF0 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing technology mainly has the following technical problems: lack of a tacrolimus individualized drug gene detection kit for the Chinese population with high detection sensitivity and high accuracy

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Gene detection kit for detecting individual drug use of tacrolimus drug
  • Gene detection kit for detecting individual drug use of tacrolimus drug
  • Gene detection kit for detecting individual drug use of tacrolimus drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] A gene detection kit for detecting individualized drug use of tacrolimus, comprising the following components:

[0026] (1) CYP3A5*3 (-253-1A>G) and POR (1508C>T) site amplification and sequencing primers 2 pairs, each primer 1OD, primer sequence see Table 1;

[0027]

[0028] (2) PCR amplification reagent: 15 μL of Taq enzyme mixture (dNTP, 10×PCR reaction buffer, MgCl2);

[0029] (3) PCR product purification reagent: 3.2 μL SAP enzyme mixture (SAP enzyme, ExoI enzyme, deionized water);

[0030] (4) Sequencing reagents: 4.4 μL BigDye mix (Bigdye, 5×seq), 5 μL EDTA, 60 μL ethanol solution (100%), 200 μL ethanol solution (70%), 16 μL Hi-Di.

[0031] This kit is stored at -20°C, and repeated freezing and thawing should be avoided as much as possible.

Embodiment 2

[0033] A method for using a gene detection kit for detecting tacrolimus drug individualized drug use, comprising the following steps:

[0034] (1) extract the genomic DNA of the sample;

[0035] (2) PCR amplification of CYP3A5*3 (-253-1A>G) and POR (1508C>T) loci

[0036] For CYP3A5*3 (-253-1A>G) and POR (1508C>T) sites, design primers according to Table 1 for PCR amplification. Each reaction system has a total volume of 16 μL, including 7.5 μL of Taq enzyme mixture (dNTP, 10×PCR reaction buffer, MgCl2), 5.5 μL of deionized water, primer pair (10uM) 1 μL, genomic DNA (100ng / ul) 1ul ;The reaction condition is 95°C for 15min, 14 cycles (-0.5°C / cycle) of 94°C for 30sec, 63°C for 30sec, 72°C for 45sec; then 30 cycles of 94°C for 30sec, 56°C for 30sec, 72°C for 45sec, Finally, 72°C for 10min;

[0037] (3) Purification of PCR products

[0038] Each reaction has a total volume of 5.6 μL. 4 μL of PCR product was added, and then 1.6 μL of SAP enzyme mixture (SAP enzyme, ExoI enzym...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a gene detection kit for detecting individualized drug use of a tacrolimus drug. The kit comprises the following components including two pairs of PCR amplification and sequencing primers for CYP3A5*3 (-253-1A>G) and POR (1508C>T) site regions, a PCR amplification reagent, a PCR product purification reagent and a DNA sequencing reagent. According to the sequencing primers for the CYP3A5*3 (-253-1A>G) site, the forward primer is TGCCCTTGCAGCATTTAG, and the reverse primer is ACACCCAGGAAGCCAGAC; according to the sequencing primers for the POR (1508C>T) site, the forward primer is GGTGGTTGTGGAGTACGAGA, and the reverse primer is CGCTCCTGGATGAAGCCTAT. The kit is used for genotyping detection of the CYP3A5*3 (-253-1A>G) and POR (1508C>T). By applying the kit in detection, the individual drug metabolism type including a normal metabolite and a slow metabolism type can be identified, the drug dosage is adjusted, the drug efficacy is improved, the toxic and side effects ofthe drug are reduced, and individualized treatment is realized.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a gene detection kit for detecting individualized drug use of tacrolimus. Background technique [0002] The calcineurin inhibitor tacrolimus is a macrolide antibiotic isolated from the metabolites of Streptomyces and was used clinically in 1989. It mainly inhibits the activation process of T cells by specifically inhibiting the activity of neural calcineurin. At the same time, it also produces T cell-dependent antibodies that cyclosporine does not have, and blocks the transcription of cytokines, presenting a double effect. Immunosuppressive effect. Tacrolimus is mainly used for graft rejection after liver or kidney transplantation. However, due to the narrow therapeutic window of tacrolimus and the large individual differences in pharmacokinetics, it is administered according to the conventional dose, and some patients may suffer from "insufficient immunosuppression" due to too low...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883
CPCC12Q1/6883C12Q2600/106C12Q2600/172
Inventor 王斐陆军姜昕唐帅男
Owner 上海佰臻生物科技有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com