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Microarray chip capable of detecting genomic DNA of various fungi and preparation method of microarray chip

A technology of microarray chips and gene chips, which is applied in the direction of biochemical equipment and methods, measurement/testing of microorganisms, etc., can solve the problems that are difficult to meet the batch chip production or chip application, and achieve the reduction of steric hindrance effect and low chip cost , The effect of simple and easy reaction

Pending Publication Date: 2020-03-06
BEIJING BEIER BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At the same time, most of the existing chips are used for scientific research, and are only suitable for operating a small number of chips or chip samples, which is difficult to meet the needs of mass chip production or chip applications.

Method used

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  • Microarray chip capable of detecting genomic DNA of various fungi and preparation method of microarray chip
  • Microarray chip capable of detecting genomic DNA of various fungi and preparation method of microarray chip
  • Microarray chip capable of detecting genomic DNA of various fungi and preparation method of microarray chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080] 1. Preparation of aldehyde-modified microarray chip carrier

[0081] In this embodiment, a commercially available conventional silicon dioxide glass slide is used as the chip solid-phase carrier body of the microarray chip, which is an ordinary glass slide for microscope inspection. The glass slide undergoes: cleaning, acid and alkali treatment, surface silanization treatment, aldehyde treatment, and cleaning to form a microarray chip carrier with aldehyde groups on the surface. Concrete preparation process is as follows:

[0082] Ordinary glass slides (76mm×25mm×1.2mm) were washed 3 times with deionized water and dried to remove water-soluble impurities adsorbed on the surface of the glass slides.

[0083] Soak the glass slide in chromic acid washing solution overnight, rinse it with deionized water after taking it out, and dry it in the air to further remove impurities such as organic matter on the surface of the glass slide.

[0084] The slides were soaked overnigh...

Embodiment 2

[0157] Example 2 Example of detection results of 6 kinds of pathogenic bacteria chips

[0158] 1. The specific method in Example 1 was used to complete the preparation of the formaldehyde-modified microarray chip carrier.

[0159]2. Design and synthesis of long-chain probes modified by amination. In this embodiment, 6 kinds of common pathogenic bacteria are detected. These 6 kinds of pathogenic bacteria are based on the summary of the incidence of different clinical fungi over the years. There may be certain differences with the total number and types of fungi detected by other inventions or methods. cross, but not exactly coincident. The detected 6 kinds of common pathogenic bacteria and their clinically possible associated symptoms, as well as the long-chain probes of C12-dT (n=10) in this method for 6 kinds of fungi, the unmodified DNA probe sequence ontology is as follows Show.

[0160]

[0161]

[0162] 3. Preparation of Microarray Chip

[0163] Prepare the dete...

Embodiment 3

[0206] On the aldylated silica glass slide, a 4×9 microarray was used to prepare a fungal chip according to the method in Example 1, and the Candida albicans probe, the positive internal control probe of the chip, and the universal fungal probe were applied. at the corresponding position on the microarray.

[0207] Candida albicans probes adopt three forms, I is a conventional 5'amino-dT(n=10)-oligonucleotide chain probe, II is a 5'amino-C12-oligonucleotide chain probe, III is the 5'amino-C12-dT (n=10) modified Candida albicans-specific probe designed in the present invention. Specifically for Candida albicans specific probes:

[0208] I: 5-NH 3 -dT(n=10)-GCATGCTGCTCTCTCGGG;

[0209] II: 5-NH 3 -C12-GCATGCTGCTCTCTCGGG;

[0210] III: 5-NH 3 -C12-dT (n=10)-GCATGCTGCTCTCTCGGG.

[0211] Spotting and fixing three different probes respectively, adding biotin-labeled Candida albicans long-fragment PCR amplification product (1500bp, which used common upstream primer ATCAATAAGFG...

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Abstract

The invention relates to a microarray chip capable of detecting genomic DNA of various fungi and a preparation method of the microarray chip. The microarray chip comprises a modified chip solid-phasecarrier, and a modified gene chip probe, wherein the modified gene chip probe comprises the following structures according to the direction of 5'-3': a 5' modified group-Cn-(dT) m-oligonucleotide chain 3', oligodeoxynucleotide is selected from nucleotide sequences of genomic DNA specific regions of two or more fungi of Candida albicans, Candida glabrata, Candida parapsilosis, Aspergillus, Candidatropicalis and Cryptococcus, or complementary sequences thereof. According to the embodiment of the invention, the modified gene chip probe in the microarray chip effectively reduces the steric hindrance effect of the probe and a PCR product during specific binding, the combination of the two modification modes plays a synergistic role, and the genomic DNA of various fungi can be well detected.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a microarray chip capable of detecting multiple fungal genome DNAs and a preparation method thereof. Background technique [0002] Fungal infection is the most common and serious clinical infectious disease, such as: central nervous system, respiratory system, blood system and other fungal infections have high mortality and disability rates, which seriously threaten the lives of patients. [0003] With the development of molecular biotechnology, the wide application of nucleic acid hybridization and chip detection technology, gene chip detection technology can be used to realize DNA detection. Gene chip detection technology can arrange different probes on a solid phase carrier to form a microarray of probes, using double-stranded DNA molecules to achieve denaturation-refolding characteristics under certain conditions, and the probes specifically bind to the DNA molecules to be tested...

Claims

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Application Information

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IPC IPC(8): C12Q1/6837C12Q1/6895C12Q1/04
CPCC12Q1/6837C12Q1/6895C12Q2600/166
Inventor 樊高君王玲闫宝山于大为邵育晓徐悦刘瑞鑫
Owner BEIJING BEIER BIOENG
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