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Microarray chip capable of detecting genome DNA (deoxyribonucleic acid) of multiple bacteria and preparation method of microarray chip

A microarray chip and gene chip technology, applied in the biological field, can solve the problems that are difficult to satisfy batch chip production or chip application, and achieve the effects of reducing steric hindrance effect, low chip cost, and simple and easy reaction

Inactive Publication Date: 2020-03-27
BEIJING BEIER BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 genome DNA (deoxyribonucleic acid) of multiple bacteria and preparation method of microarray chip
  • Microarray chip capable of detecting genome DNA (deoxyribonucleic acid) of multiple bacteria and preparation method of microarray chip
  • Microarray chip capable of detecting genome DNA (deoxyribonucleic acid) of multiple bacteria and preparation method of microarray chip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] 1. Preparation of aldehyde-modified microarray chip carrier

[0080] 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:

[0081] 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.

[0082] 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.

[0083] The slides were soaked overnigh...

Embodiment 2

[0156] Example 2 Example of detection results of 16 kinds of pathogenic bacteria chips

[0157] 1. The specific method in Example 1 was used to complete the preparation of the formaldehyde-modified microarray chip carrier.

[0158] 2. Design and synthesis of long-chain probes modified by amination. In this embodiment, 16 kinds of common pathogenic bacteria are detected. These 16 kinds of pathogenic bacteria are based on the summary of the incidence of different clinical bacteria over the years. There may be certain differences with the total number and types of bacteria detected by other inventions or methods. cross, but not exactly coincident. The detected 16 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 16 kinds of bacteria, the unmodified DNA probe sequence ontology is as follows Show.

[0159]

[0160]

[0161] 3. Preparation of Microarray Chip

[0162] A...

Embodiment 3

[0240] On the aldylated silica glass slide, adopt 8 * 12 microarray form to prepare bacterial chip according to the method in embodiment 2, enterobacteria faecalis probe, positive quality control probe of the chip, bacterial universal probe, leather Gram stain positive probes and Gram stain negative probes are spotted on the corresponding microarray positions.

[0241] The Enterobacter faecalis probe adopts two forms respectively, I is the conventional 5'amino-dT(n=10)-oligonucleotide chain probe, II is the 5'amino-C 12-dT(n=10) designed by the present invention =10)-oligonucleotide chain probe, specifically Enterobacter faecalis specific probe I:5-NH 3 -dT(n=10)-TCGTTAGTACATGAACGTCCCCTG. Enterobacter faecalis specific probe II: 5-NH 3 -C12-dT (n=10)-TCGTTAGTACATGAACGTCCCCTG.

[0242] Two different Enterobacter faecalis probes were spotted and fixed respectively, and the biotin-labeled Enterobacter faecalis 16S long-chain PCR amplification product (1500bp, which was amplifi...

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Abstract

The embodiment of the invention relates to a microarray chip capable of detecting genome DNA (deoxyribonucleic acid) of multiple bacteria and a preparation method of the microarray chip. The microarray chip comprises a chip solid-phase vector body and a modified gene chip probe, wherein the modified gene chip probe comprises the following structure in a 5'-3' direction: 5' modied group-Cn-(dT)m-oligonucleotide chain 3'; and the oligonucleotide chain is selected from nucleotide sequences or complementary sequences of the nucleotide sequences in genome DNA specific areas of two or more than twobacteria such as acinetobacter baumannii, klebsiella pneumoniae, escherichia coli and pseudomonas aeruginosa. Through the modified gene chip probe in the microarray chip provided by the embodiment ofthe invention, the steric-hinerance effect of the probe with a PCR (polymerase chain reaction) product in specific combination can be effectively reduced, combination of two modification modes plays arole of synergism, and the genome DNA of multiple bacteria 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 bacterial genome DNAs and a preparation method thereof. Background technique [0002] Bacterial infection is the most common and serious clinical infectious disease, such as: central nervous system, respiratory system, blood system and other bacterial 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6806C12Q1/6837C12Q1/14C12Q1/10C12Q1/04C12R1/22C12R1/385C12R1/445C12R1/46C12R1/21C12R1/36C12R1/32C12R1/44C12R1/01
CPCC12Q1/689C12Q1/6806C12Q1/6837C12Q2531/113
Inventor 徐悦王玲闫宝山于大为邵育晓刘瑞鑫樊高君
Owner BEIJING BEIER BIOENG
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