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Method of testing PML/RAR alpha genes through electrochemical biosensor of carbon dot @ graphene oxide composite material

A biosensor and composite material technology, applied in the field of electrochemical DNA biosensor detection of PML/RARα fusion gene, can solve the problems of complicated RT-PCR operation, time-consuming and laborious chromosome analysis, complicated method, etc., and achieve good conductivity and DNA loading capacity. , Conducive to promotion and use, the effect of simple operation process

Inactive Publication Date: 2016-02-17
FUJIAN MEDICAL UNIV
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Problems solved by technology

[0009] At present, the clinical diagnosis methods of APL mainly include hybridization techniques such as chromosome analysis, Southern-Blot, real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH), but these methods have certain limitations. : Chromosome analysis is time-consuming and labor-intensive, with low sensitivity; Southern-Blot is extremely insensitive to the detection of low-copy gene sequences, not only the method is complex, the detection cycle is long, the price is expensive, and there is radioactive contamination, which limits the wide application in clinical practice; RT-PCR The operation is cumbersome and expensive; FISH needs to use a complex fluorescent color system, and its relative sensitivity is not high, making it difficult to carry out large-scale clinical application

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  • Method of testing PML/RAR alpha genes through electrochemical biosensor of carbon dot @ graphene oxide composite material
  • Method of testing PML/RAR alpha genes through electrochemical biosensor of carbon dot @ graphene oxide composite material
  • Method of testing PML/RAR alpha genes through electrochemical biosensor of carbon dot @ graphene oxide composite material

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Embodiment 1

[0038] The design steps of the PML / RARα fusion gene probe are as follows:

[0039] (1) Search the PML / RARα fusion gene sequence from the NCBI gene database, select one of them as a synthetic probe, the length of the probe is 24 bases, and use the corresponding sequence as the PML / RARα fusion gene sequence after homologous comparison. The specific sequence of RARα was synthesized, and the capture probe (5'-NH 2 -GGTCTCAATGGCTGCCTCCCCG-3'), target DNA sequence (5'-CGGGGAGGCAGCCATTGAGACC-3').

Embodiment 2

[0041] The electrochemical biosensor preparation steps of carbon dot graphene oxide composites are as follows ：

[0042] (1) Glassy carbon electrode (GCE) in the order of 1.0, 0.3, 0.05μm Al 2 o 3 Grinding in powder, polishing. Then put them into 1:1 nitric acid solution (that is, HNO with a concentration of 16mol / L 3 The volume ratio with water is 1:1), ultrasonication in absolute ethanol and double distilled water and blown dry with nitrogen, set aside.

[0043] (2) The characterization results of carbon dots (C-dots for short) are shown in figure 2 . Such as figure 2 As shown in A, the prepared carbon dots are approximately spherical. According to the statistics of the particle size of the carbon dots according to the TEM data, the particle size is about 0.97~2.18nm (see figure 2 in B). From high-resolution transmission electron microscopy of carbon dots such as figure 2 C in and figure 2 As shown in D, the prepared carbon dots have obvious lattice characteri...

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Abstract

The invention discloses a method of testing PML / RAR alpha genes through an electrochemical biosensor of a carbon dot @ graphene oxide composite material. The method comprises the steps that (1) a specific probe is designed according to a gene segment to be tested, wherein the PML / RAR alpha fusion genes of acute promyelocytic leukemia are used as an example; (2) capture probes are self-assembled on the C-dots @ GO / GCE surface through amido bonds, and after hybridization of a DNA electrochemical sensor formed with methylene blue (MB) as the electrochemical hybridization indicator with a solution to be tested, whether the solution to be tested contains the PML / RAR alpha fusion genes or not is judged according to difference of electrical signals. The C-dots @ GO nanocomposite prepared through the method has good electrical conductivity, and abundant carboxyl on the GO surface can increase the number of the capture probes; the DNA sensor formed on such basis has the advantages of good selectivity and high sensitivity.

Description

technical field [0001] The invention relates to a method for detecting a PML / RARα fusion gene by an electrochemical DNA biosensor based on a carbon-dot graphene oxide composite material, and belongs to the technical field of biosensing. Background technique [0002] Nanotechnology is mainly a technology aimed at the molecular world with a size between 1nm and 100nm. This size is in the transition zone between the microscopic world represented by atoms and molecules and the macroscopic object. The system based on this size is neither a typical microscopic system nor a typical macroscopic system, so it has unique physical and chemical properties, such as surface effects, Micro-scale effects, quantum effects, and macro-quantum tunneling effects, etc., reflect the superior performance that conventional materials do not have. [0003] A biosensor is composed of a molecular recognition part (sensitive element) and a conversion part (transducer). The molecular recognition part is ...

Claims

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Application Information

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IPC IPC(8): G01N27/30
CPCG01N27/308
Inventor 雷云刘爱林林新华陈伟王鹏邓娅妮郑珍妮
Owner FUJIAN MEDICAL UNIV
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