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A nucleic acid aptamer fluorescent test strip and its preparation method and application

A nucleic acid aptamer and test strip technology, which is applied in the direction of instruments, measuring devices, scientific instruments, etc., can solve the problems of inability to achieve sensitive and accurate detection, inability of aptamers to detect line binding, inability to achieve precise detection of targets, etc. Achieve the effect of sensitive, accurate and reliable measurement results, simple structure, and improved comparability

Active Publication Date: 2020-03-27
INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are not many colloidal gold or fluorescent test strips based on nucleic acid aptamers. The main reason is that different aptamer sequences have different lengths. It is greater than the affinity of the complex formed by the aptamer and the target, so the aptamer that has bound to the target still dissociates at the detection line, and the accurate detection of the target cannot be achieved
Although the fixed antigen on the detection line makes the competitiveness consistent, for many small molecule targets, the combination with the coupling protein limits the binding to the aptamer, making the aptamer unable to bind at the detection line, and cannot achieve sensitive and accurate detection

Method used

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  • A nucleic acid aptamer fluorescent test strip and its preparation method and application
  • A nucleic acid aptamer fluorescent test strip and its preparation method and application
  • A nucleic acid aptamer fluorescent test strip and its preparation method and application

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preparation example Construction

[0035] A preparation method of nucleic acid aptamer fluorescent test strip comprises:

[0036]Preparation method of nanomaterial-modified nanomaterial mat: select cellulose film as the raw material for preparing nanomaterial mat, spray nanomaterial suspension solution onto the cellulose film, dry at 110-130°C for 7-9 hours, and remove glass fiber Moisture on the plain film was obtained to obtain a cellulose film modified by nanomaterials. The nanomaterial pad can be obtained by cutting according to needs.

[0037] Preparation method of nanomaterial pads adsorbed with fluorescently labeled aptamers: add fluorescently labeled aptamers and / or fluorescently labeled aptamers with linking sequences and free fluorescein to the nanomaterials, and incubate for 15 After ~25 minutes, dry under the condition of 35-40° C. to remove the moisture of the nanomaterials. Preferably, the fluorescein can be fluorescein isothiocyanate (FITC), phycoerythrin (PE), anthocyanin 5 (Cy5), carboxyfluor...

Embodiment 1

[0048] Please refer to figure 1 , the present invention provides a nucleic acid aptamer fluorescent test strip 100, including a bottom plate 160, a sample pad 110, a nanomaterial pad 120, a reaction membrane 130, and a water-absorbing pad 140, a sample pad 110, a nanomaterial pad 120, and a reaction membrane 130 And the ends of the water-absorbing pad 140 are sequentially connected to form the detection layer 150 , and the detection layer 150 is connected to the bottom plate 160 .

[0049] In the embodiment of the present invention, the nucleic acid aptamer fluorescent test strip 100 can be a conventional strip shape, and the sample pad 110, the nanomaterial pad 120, the reaction membrane 130 and the water-absorbing pad 140 are all corresponding strip shapes, In other embodiments, the nucleic acid aptamer fluorescent test strip 100 can be circular or other shapes, and the shapes of the sample pad 110, the nanomaterial pad 120, the reaction membrane 130, and the water-absorbing...

Embodiment 2

[0052] This embodiment provides a sulfadimethoxine nucleic acid aptamer test strip, its preparation method and its detection and application.

[0053] One, the preparation method of sulfadimethoxine nucleic acid aptamer test strip comprises:

[0054] Select glass cellulose film as the raw material for preparing nanomaterial mats, spray the graphene oxide suspension solution on the glass cellulose film, dry at 120°C for 8 hours, remove the moisture on the glass cellulose film, and obtain the modified graphene oxide Sexual fiberglass. Cutting to get nanomaterials.

[0055] Add the Cy5 fluorescein-labeled sulfadimethoxine aptamer (Cy5-GAGGG-CAACGAGTGTTTATAGATTTTTTTTTTTTTTTTT) containing the linker sequence polyT15 dropwise onto the nanomaterial pad, incubate for 20 minutes, and dry at 37°C to remove the moisture of the nanomaterial .

[0056] A nitrocellulose membrane was chosen as the reaction membrane. The complementary sequence polyA15 (Btiotin-AAAAAAAAAAAAAAAA) of the bio...

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Abstract

The invention provides a nucleic acid aptamer fluorescence test strip, a method for preparing the same and application of the nucleic acid aptamer fluorescence test strip, and relates to the technicalfield of biological detection. The nucleic acid aptamer fluorescence test strip comprises a bottom plate, a sample pad, a nanometer material pad, a reaction film and a water absorption pad. The reaction film is provided with a detection line, and the detection line is arranged at the end, which is close to the nanometer material pad, of the reaction film; nucleic acid aptamer with fluorescein labels and / or nucleic acid aptamer with fluorescence labels and linker sequences and free fluorescein are attached to the nanometer material pad, and complementary strains of nucleic acid aptamer or complementary strains of linker sequences of nucleic acid aptamer are fixed to the detection line. The method for preparing the nucleic acid aptamer fluorescence test strip includes sequentially adheringthe sample pad, the nanometer material pad, the reaction film and the water absorption pad on the same side of the bottom plate. The nucleic acid aptamer fluorescence test strip, the method and the application have the advantages that the nucleic acid aptamer fluorescence test strip is simple in structure, sensitive in detection and high in accuracy; the method is easy to implement and high in flexibility.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a nucleic acid aptamer fluorescent test strip and its preparation method and application. Background technique [0002] As a simple and fast analytical tool, test strips have been widely used in clinical diagnosis, food safety and environmental monitoring. At present, there are mainly two detection methods of colloidal gold and fluorescence test strips. Among them, the colloidal gold test strips display the results through color reaction, which has the advantage of being visible to the naked eye without the need for other instruments; but its disadvantage is low sensitivity, and can only be tested by comparison The color depth achieves the purpose of semi-quantitative. Compared with colloidal gold test strips, fluorescent test strips are highly sensitive, and can be used with a portable fluorescence detector to accurately quantify the target in the sample, so it has...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/533G01N33/559
CPCG01N33/533G01N33/559
Inventor 陈爱亮张桂兰朱超杨曙明
Owner INST OF QUALITY STANDARD & TESTING TECH FOR AGRO PROD OF CAAS
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