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H2 receptor blocking drug efficacy related gene SLC22A1 polymorphism detection kit

A technology of polymorphism detection and receptor blocking, applied in the biological field, achieves short time-consuming, convenient use, and strong specificity

Inactive Publication Date: 2018-07-24
上海美丽人生医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a lack of rapid and effective methods for detecting the relevant mutation points of SLC22A1 mutation heterozygotes and mutation homozygotes

Method used

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  • H2 receptor blocking drug efficacy related gene SLC22A1 polymorphism detection kit
  • H2 receptor blocking drug efficacy related gene SLC22A1 polymorphism detection kit
  • H2 receptor blocking drug efficacy related gene SLC22A1 polymorphism detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] A primer composition for detecting the SLC22A1 polymorphism of an H2 receptor blocking drug efficacy-related gene, including a C1022T detection primer and an internal control gene detection primer, the sequences of which are shown in Table 1 and Table 2 below.

[0054] Table 1 Sequences of detection primers for SLC22A1 gene polymorphisms (ie, C1022T detection primers)

[0055] primer name

Base sequence (5′→3′ direction)

P1-FW1 (C1022T detection primer 1)

5′-GCAGACCTGTTCCGCACGTC-3

P1-FW2 (C1022T detection primer 2)

5′-CAGACCTG TTCCGCACGAT-3′

P1-R (C1022T detection primer three)

5′-CACTCACAACTATATTACA-3′

[0056] Table 2 Sequences of primers for internal control gene detection

[0057] primer name

Embodiment 2

[0059] like figure 1 As shown, the kit containing the primer composition as described in Example 1 includes: a box body 1 with a box lid, a baffle plate 3, two sponges 2, two test tubes containing distilled water 4, two containing PCR Test tube for reaction master mix 5, one test tube containing positive quality control SLC22A1 gene C1022T mutant plasmid 6, one test tube containing positive quality control SLC22A1 gene C1022T wild plasmid 7, two test tubes containing internal control gene detection primers 8, three test tubes Tube 9 with primers for SLC22A1 gene detection. Wherein, the baffle 3 is placed in the box body 1, the box body 1 is divided into two parts, the two sponges 2 are respectively placed in the box body 1 on both sides of the baffle plate 3, the sponge 2 is punched with container holes 10, and the two One test tube containing distilled water 4, two test tubes containing PCR reaction premix 5 placed in the container hole 10 of sponge 2 on one side of baffle 3...

Embodiment 3

[0062] The operation method of the test kit of above-mentioned embodiment 2 is:

[0063] (1) Sample processing and nucleic acid extraction

[0064] The samples were treated with commercial DNA extraction kits, and the concentration and purity of the extracted DNA should be determined with a UV spectrophotometer. 260 / OD 280 The value should be between 1.8 and 2.0, and the concentration should be between 5 and 50 ng / μL. Those with unqualified DNA quality should not be used for detection. It is recommended to re-extract nucleic acid if the DNA quality is lower than 5 ng / μL, and appropriate if it is higher than 50 ng / μL. Dilute to the specified concentration range. It is recommended to test the extracted DNA immediately. Otherwise, please store it below -20°C for no longer than 6 months. The kit quality control materials (ie, various plasmids) were thawed at room temperature before use, vortexed for 10 seconds, and centrifuged at 2000 rpm for 15 seconds before use.

[0065] (2...

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Abstract

The invention relates to an H2 receptor blocking drug efficacy related gene SLC22A1 polymorphism detection kit, comprising a test tube separately provided with each C1022T detection primer and each internal control gene detection primer, wherein the C1022T detection primer comprises the following sequence components: C1022T detection primer 1: 5'-GCAGACCTGT TCCGCACGTC-3'; C1022T detection primer 2: 5'-CAGACCTGTT CCGCACGAT-3'; C1022T detection primer 3: 5'-CACTCACAAC TATATTACA-3'; the internal control gene detection primer comprises the following sequence components: internal control primer 1:5'-AGCAAGCAGG AGTATGACG-3'; internal control primer 2: 5'-GAAAGGGTGT AACGCAACT-3'. Compared with the prior art, the kit has the advantages of high sensitivity, high specificity and short time consumption when the SLC22A1 gene mutation of samples such as fresh blood is detected.

Description

technical field [0001] The invention belongs to the technical field of the field of biotechnology, and relates to a primer composition, a kit and a method for detecting the SLC22A1 polymorphism of an H2 receptor blocking drug efficacy-related gene. Background technique [0002] H2 receptor blocking drugs include cimetidine, ranitidine, famotidine, etc., which have the effect of resisting histamine to stimulate the secretion of hydrochloric acid from the gastric cell wall, and are widely used in peptic ulcer. Oral absorption of ranitidine is rapid but incomplete, with a bioavailability of only 50%. In the process of ranitidine transport, organic cation transporter (OCT1) plays a key role. SLC22A1 is the gene encoding OCT1, and its 1022C>T gene polymorphism can significantly affect the pharmacokinetics of ranitidine in humans. The study found that the bioavailability of ranitidine was more efficient in populations carrying SLC22A1 mutation heterozygotes and mutation homoz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/106C12Q2600/156
Inventor 田晓丽
Owner 上海美丽人生医疗科技有限公司
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