Gene detection chip of OATP1B1 major gene mutation

Abstract

The invention relates to a gene detection chip, in particular to a gene detection chip used for detecting common major gene mutation of organic anion transferred polypeptide 1B1 (OATP1B1 in short), which is closely related to reactiveness of medicines used for curing high blood pressure, malignant tumor, 2 type diabetes mellitus, hypercholesterolemia. Through selecting 8 mutational sites and designing out corresponding probes, the invention realizes the detection on the mutational sites and is capable of providing information of the mutation sites, thereby providing foundation for medication guiding.

Description

technical field

[0001] The invention belongs to the field of gene analysis and detection, and in particular relates to a gene detection chip, more specifically, it is used to detect the reactivity of drugs closely related to hypertension, malignant tumors, type 2 diabetes and hypercholesterolemia. Gene detection chip for common important gene mutations of organic anion transporting polypeptide 1B1 (OATP1B1 for short). Background technique

[0002] According to statistics, the number of patients with hypertension and diabetes in my country is about 130 million and 20 million respectively, the incidence of dyslipidemia in adults is about 18.6%, and the number of patients with malignant tumors is about 1.8 to 2 million. At present and for a long time in the future, drugs It will still be the main method for the treatment of the above major diseases. The liver is the main organ for the metabolism and detoxification of drugs and exogenous substances in the human body. Organic ani...

Examples

Embodiment 1

[0041] Example 1: Preparation of a chip detection system for OATP1B1 common mutation gene sites

[0042] 1. Detailed preparation process

[0043] 1.1. Substrate treatment: The substrate is made of glass medium, and the surface treatment method is aldehyde modification. The specific steps are:

[0044] 1. Selection of blank super-flat film: choose 76mm×25mm×1mm super-flat film, the error of its length and width does not exceed 0.2mm, the error of thickness does not exceed 0.1mm, no damage, no scratches on the surface;

[0045] 2. Ultra-flat sheet pretreatment: soak the ultra-flat sheet in the newly prepared potassium dichromate lotion, leave it for 7 days, wash it with deionized water, and dry it;

[0046] 3. Amination of ultra-flat slides: Prepare ethanol solutions of aminosilane with a concentration of 2%, immerse the slides in the above solutions for 15 minutes, take them out, wash them with deionized water, and dry them in the air;

[0047] 4. Aldehylation of super-flat ...

Embodiment 3

[0096] [Example 3] Comparison of hybridization effects of probes with different base numbers

[0097] In the present invention, when the probes of the sequences shown in Table 3 are actually used, a continuous 14-25 base sequence can be selected to synthesize the probe. The following experiments are designed to prove that the difference in the number of bases above does not affect the sensitivity of the results and specificity.

[0098] For the -11187G>A mutation of the OATP1B1 gene, the following mutant and normal probes of different lengths were synthesized:

[0099] Wild-type probe I (base sequence from 19th to 34th in SEQ ID NO.1, 15 bases in total)

[0100] 5'-GTATACA GTAAAAG-3'

[0101] Mutant probe I (19th to 34th base sequence of SEQ ID NO.3, 15 bases in total)

[0102] 5'-GTATACA GTAAAAG-3'

[0103] Wild-type probe II (17th to 36th base sequence of SEQ ID NO.1, 19 bases in total)

[0104] 5'-ATGTATACA GTAAAAGTG-3'

[0105] Mutant probe II (17th to 36th bas...

technical field

[0001] The invention belongs to the field of gene analysis and detection, and in particular relates to a gene detection chip, more specifically, it is used to detect the reactivity of drugs closely related to hypertension, malignant tumors, type 2 diabetes and hypercholesterolemia. Gene detection chip for common important gene mutations of organic anion transporting polypeptide 1B1 (OATP1B1 for short). Background technique

[0002] According to statistics, the number of patients with hypertension and diabetes in my country is about 130 million and 20 million respectively, the incidence of dyslipidemia in adults is about 18.6%, and the number of patients with malignant tumors is about 1.8 to 2 million. At present and for a long time in the future, drugs It will still be the main method for the treatment of the above major diseases. The liver is the main organ for the metabolism and detoxification of drugs and exogenous substances in the human body. Organic ani...

Examples

Embodiment 1

[0041] Example 1: Preparation of a chip detection system for OATP1B1 common mutation gene sites

[0042] 1. Detailed preparation process

[0043] 1.1. Substrate treatment: The substrate is made of glass medium, and the surface treatment method is aldehyde modification. The specific steps are:

[0044] 1. Selection of blank super-flat film: choose 76mm×25mm×1mm super-flat film, the error of its length and width does not exceed 0.2mm, the error of thickness does not exceed 0.1mm, no damage, no scratches on the surface;

[0045] 2. Ultra-flat sheet pretreatment: soak the ultra-flat sheet in the newly prepared potassium dichromate lotion, leave it for 7 days, wash it with deionized water, and dry it;

[0046] 3. Amination of ultra-flat slides: Prepare ethanol solutions of aminosilane with a concentration of 2%, immerse the slides in the above solutions for 15 minutes, take them out, wash them with deionized water, and dry them in the air;

[0047] 4. Aldehylation of super-flat ...

Embodiment 3

[0096] [Example 3] Comparison of hybridization effects of probes with different base numbers

[0097] In the present invention, when the probes of the sequences shown in Table 3 are actually used, a continuous 14-25 base sequence can be selected to synthesize the probe. The following experiments are designed to prove that the difference in the number of bases above does not affect the sensitivity of the results and specificity.

[0098] For the -11187G>A mutation of the OATP1B1 gene, the following mutant and normal probes of different lengths were synthesized:

[0099] Wild-type probe I (base sequence from 19th to 34th in SEQ ID NO.1, 15 bases in total)

[0100] 5'-GTATACA GTAAAAG-3'

[0101] Mutant probe I (19th to 34th base sequence of SEQ ID NO.3, 15 bases in total)

[0102] 5'-GTATACA GTAAAAG-3'

[0103] Wild-type probe II (17th to 36th base sequence of SEQ ID NO.1, 19 bases in total)

[0104] 5'-ATGTATACA GTAAAAGTG-3'

[0105] Mutant probe II (17th to 36th bas...

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