Gene detection chip of common mutations of diphosphate glucuronyl transferase 1A and use method thereof

A technology of gene chip and mutant gene, applied in the field of gene detection chip, can solve the problems such as the disclosure of the technical scheme of ungene chip, and achieve the effect of avoiding difference in results, less demand, and reducing human error.

Active Publication Date: 2014-04-09
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] At present, there is no gene chip and its technical solution for comprehensive and systematic detection of important gene mutations in all the first exons and regulatory regions of the above-mentioned UGT1A

Method used

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  • Gene detection chip of common mutations of diphosphate glucuronyl transferase 1A and use method thereof
  • Gene detection chip of common mutations of diphosphate glucuronyl transferase 1A and use method thereof
  • Gene detection chip of common mutations of diphosphate glucuronyl transferase 1A and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1: Preparation of a microarray detection system for common mutations in UGT1A

[0043] 1. Detailed preparation process

[0044] 1.1. Substrate treatment: The substrate is made of glass medium, and the surface treatment method is aldehyde modification. The specific steps are:

[0045] 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;

[0046] 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;

[0047] 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;

[0048] 4. Aldehylation of super-flat she...

Embodiment 3

[0105] [Example 3] Comparison of hybridization effects of probes with different base numbers

[0106] 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.

[0107] The following mutant and normal probes of different lengths were synthesized for the UGT1A6T19G mutation:

[0108] Wild-type probe I (15th to 29th base sequence of SEQ ID NO.21, 15 bases in total)

[0109] 5'-TCCTTCGCTCATTTC-3'

[0110] Mutant probe I (15th to 29th base sequence of SEQ ID NO.23, 15 bases in total)

[0111] 5'-TCCTTCGCGCATTTC-3'

[0112] Wild-type probe II (base sequence from 13th to 32nd in SEQ ID NO.21, 19 bases in total)

[0113] 5'-CCTCCTTCGCTCATTTCAG-3'

[0114] Mutant probe II (13th to 32nd base sequence of SEQ ID NO...

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Abstract

The invention relates to a gene detection chip and in particular relates to a gene detection chip which is utilized to detect the common and important mutations of diphosphate glucuronyl transferase 1A (UGT1A) which is closely related to the malignant tumors, the immunodepression and the reactivity of medicines for curing mental diseases and also related to the diagnosis of hereditary hyperbilirubinemia. Proper gene mutational sites are selected to further obtain a probe for detecting the sites and the probe is utilized to prepare the gene chip, thus the mutational sites can be detected efficiently and accurately, the relevant information can be obtained, and clinical medication and disease prevention can be guided.

Description

Technical field: [0001] The invention relates to a gene detection chip, more specifically, it is used to detect uridine diphosphate glucose which is closely related to the reactivity of malignant tumors, immunosuppression, and mental disease treatment drugs, and the diagnosis of hereditary hyperbilirubinemia A gene detection chip for common and important gene mutations of aldolyl transferase 1A (UGT1A for short). Background technique: [0002] The mortality rate of malignant tumors ranks first in my country's overall mortality rate, about 180 / 100,000 people, and is the number one killer threatening people's health. And the incidence of mental illness also increases sharply with the accelerated pace of life. According to statistics, the incidence of mental illness in big cities has reached about 13%. The number of hypertensive patients in my country is as high as 130 million. At present and for a long time in the future, drugs will still be the main means of treatment for t...

Claims

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Application Information

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