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Kit and method for detecting OATP1B1 genetic polymorphism by use of pyrosequencing technique

A pyrosequencing method and gene polymorphism technology, applied in the field of molecular biology, can solve the problems of decreased lipid-lowering efficacy, increased toxic and side effects, and increased blood drug concentration, achieving the effect of small sample size and easy operation

Inactive Publication Date: 2014-07-09
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For statins that are mainly transported by OATP1B1, when SLCO1B1 mutations lead to decreased function, the amount of SLCO1B1 entering the liver cells will decrease, the lipid-lowering effect will decrease, the blood drug concentration will increase, and the side effects will increase

Method used

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  • Kit and method for detecting OATP1B1 genetic polymorphism by use of pyrosequencing technique
  • Kit and method for detecting OATP1B1 genetic polymorphism by use of pyrosequencing technique
  • Kit and method for detecting OATP1B1 genetic polymorphism by use of pyrosequencing technique

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Embodiment 1

[0029] OATP1B1-pyroF (upstream primer): 5'-ACA CTC TCT TAT CTA CAT AGG TTG-3' (SEQ ID NO.2);

[0030] OATP1B1-pyroR (downstream primer): 5'-GGT ACT ATG GGA GTC TCC CCTATT-3' (SEQ ID NO.3);

[0031] Sequencing primer: 5'-CAC GAA GCA TAT TAC CCA T-3' (SEQ ID NO.4);

[0032] 1. DNA extraction

[0033] 1.1 The preparation and inspection of reagent materials before the experiment are as follows:

[0034] (1) Check the shelf life of the kit and ensure that ethanol has been added to Wash Buffer 1 and 2, and tick the corresponding mark on the bottle; (2) Isopropanol (if not available, it can be replaced by absolute ethanol) and 75% ethanol ;

[0035] (3) 1.5mL Eppendorf tubes and various pipette tips within the validity period of autoclaving.

[0036] 1.2 Take out the EDTA anticoagulant tube containing whole blood from the refrigerator at 4°C, and mix it upside down several times;

[0037] 1.3 Make a mark on the 1.5mL Eppendorf tube corresponding to the unique identification of t...

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Abstract

The invention discloses a kit and method for detecting OATP1B1 genetic polymorphism by use of the pyrosequencing technique. The kit checks the OATP1B1 genetic polymorphism which specifically refers to the rs4149056 (T>C) single nucleotide polymorphism. The kit comprises the primer shown by SEQ ID NO.2-4. The kit disclosed by the invention can realize accurate, quick and high-flux detection on the OATP1B1 genetic polymorphism so as to realize safe, reasonable and effective personalized administration of the substrate.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a kit and a method for detecting OATP1B1 gene polymorphism by pyrosequencing. Background technique [0002] The liver is the main organ for biotransformation and detoxification. There are many types of drug transporters in the liver, which play an important role in the absorption, distribution and excretion of drugs, such as multidrug resistance-associated proteins, breast cancer resistance proteins, organic anion transporters Peptide family, etc. OATP1B1 is the most important member of the organic anion transporting polypeptide family. The coding gene SLCO1B1 of OATP1B1 is not only highly polymorphic, but also has multiple functionally significant mutants. Studies have confirmed that OATP1B1 not only targets the transport of unconjugated bilirubin, bile acids, thyroid hormones and other endogenous substances, but also mediates the liver transmembrane transport of a ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 周宏灏
Owner CENT SOUTH UNIV