Uses and method for polymorphic point genetype for predicting sulfaurea drugs effect

A technology of polymorphic sites and sulfonylureas, applied in the field of medicine, can solve the problems of inability to improve the efficacy of drugs, toxic side effects, economic losses of patients, and delays in treatment timing, etc.

Inactive Publication Date: 2007-08-01
BEIJING HUAANFO BIOMEDICAL RES CENT +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This study suggests that such genetic variants may make sulfonylureas less effective
[0010] Due to individual differences in drug reactions, doctors cannot choose drugs and drug compatibility according to the i

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1: Determination of the Ser1369Ala (dsSNP ID: rs757110, TCC→GCC) polymorphism site genotype of the SUR1 gene and prediction of the effect of sulfonylureas

[0048] (1) Determination of the genotype of the rs757110 Ser1369Ala polymorphism site of the SUR1 gene:

[0049] (1) Extract the genomic DNA of the host cell:

[0050] (a) Add 30ml of erythrocyte lysate to the whole blood, shake slowly, and let stand at room temperature for 10 minutes. During this period, shake several times to completely lyse the erythrocytes;

[0051] (b) Centrifuge at 4°C and 2000 rpm for 10 minutes, remove the supernatant, break up the precipitated white blood cells on a rotary shaker, add 40ul of protease and 50ul of RNase, shake well, add 15ml of white blood cell lysate, and mix Take it out after evenly bathing in 37°C water bath for 20 minutes, and put it in cold water;

[0052] (c) Add 4ml of cold protein precipitation solution, mix well and place in -20°C refrigerator for 5 minute...

Embodiment 2

[0191] Example 2: Determining the genotype of the CT (rs2074312) polymorphism of the SUR1 gene and predicting the hypoglycemic effect of sulfonylureas

[0192] (1) Determination of the genotype of the CT (rs2074312) polymorphism site of the SUR1 gene:

[0193] (1) According to the conventional operation process, adopt the method similar to Example 1 to extract the genomic DNA of the host cell

[0194] (2) Use Taqman method to detect the genotype of the CT (rs2074312) polymorphism site of the SUR1 gene

[0195](a) Amplify the SUR1 functional gene polymorphic site and its flanking sequences with a PCR instrument, containing 10ng of genomic DNA in a 5ul PCR reaction system, and 2.5ul of Taqman 2X Universal PCR Master Mix No AmpErase UNG (composition includes: AmpliTaq Gold DNA Polymerase, dNTPs with Dutp, Passive Reference, optimized buffer), and 0.72uM forward primer, 0.72uM reverse primer and two allele-specific probes with fluorescent reporter groups each 0.16uM.

[0196] Th...

Embodiment 3

[0223] Example 3: Determination of the CT (rs1799854, 16-3c / t) polymorphism site genotype of the SUR1 gene and prediction of the hypoglycemic effect of sulfonylureas

[0224] (1) Determination of the genotype of the CT (rs1799854, 16-3c / t) polymorphism site of the SUR1 gene:

[0225] (1) Extract the genomic DNA of the host cell:

[0226] On the basis of standard operating procedures, the genomic DNA in salivary cells was extracted by salt precipitation. DNA was stored at -20°C.

[0227] (2) Use the TaqMan method to detect the CT (rs1799854, 16-3c / t) polymorphic site genotype of the SUR1 gene:

[0228] (a) Amplify the CT (rs1799854, 16-3c / t) gene polymorphic site of the SUR1 gene and its flanking sequences with a PCR machine, and use a 5ul PCR reaction system containing 10ng of genomic DNA and 2.5ul of TaqMan 2X Universal for PCR Master Mix No AmpErase UNG (composition includes: AmpliTaq Gold DNA Polymerase, dNTPs with Dutp, Passive Reference, optimized buffer), and 0.72uM f...

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PUM

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Abstract

The invention relates to the prediction of sulfonylureas drug effect by measuring polymorphism site genetype of sulfonylureas recepted gene, and the method. It predicts the effect of sulfonylureas drug in reducing blood sugar, improving and restoring Langer-hans' insula beta cell function and/ or intensifying insulin sensitive function by measuring polymorphism site genetype of sulfonylureas recepted gene; and predicts the effect of sulfonylureas drug in reducing blood sugar, improving and restoring Langer-hans' insula beta cell function and/ or intensifying insulin sensitive function by making use of polymorphism parting oligonucleotide to detect polymorphism site genetype of sulfonylureas recepted gene in biological sample. The invention can indicate doctor to choose medicine according to individual gentic difference, increase the effective rate of clinical medicine treating and safty of clinical midicine usage, reduce toxic and side effect of medicine and economical load, and reduce diabetes complication generation.

Description

technical field [0001] The present invention relates to the use and method of polymorphic site genotype prediction of the effect of sulfonylurea drugs, especially comprising at least Ser1369Ala (rs757110), C / T (rs2074312) and C / The application and method for predicting the action and effect of sulfonylurea drugs by the polymorphic site of the sulfonylurea receptor gene of T(rs1799854) belong to the field of medicine. Background technique [0002] Sulfonylurea antidiabetic drugs are insulin secretagogues that lower fasting and postprandial blood glucose. For most patients with newly diagnosed type 2 diabetes, sulfonylurea antidiabetic drugs can reduce fasting blood sugar by 50-80 mg / dl and glycosylated hemoglobin (HbAlc) by 1.0%-2.5%. Because β-cell function declines over time in patients with type 2 diabetes, sulfonylurea antidiabetic drugs are very effective in lowering blood glucose in patients with clinically newly diagnosed diabetes. Sulfonylurea antidiabetic drugs an...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/48
Inventor 徐希平任晓炜冯雁邢厚恂王燕蒋善群戴成祥
Owner BEIJING HUAANFO BIOMEDICAL RES CENT
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