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Construction method of fluorescent aptamer sensor and application of fluorescent aptamer sensor in novel coronavirus detection

An aptamer sensor and construction method technology, applied in the field of biomedicine, can solve the problems of limited use range, high cost, difficult modification, etc., and achieve the effects of improving detection sensitivity, simple and fast operation, stability and repeatability

Active Publication Date: 2022-02-01
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Biosensors for virus antigen detection use antibodies as biorecognition elements, which are costly and difficult to modify, which limits the scope of use

Method used

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  • Construction method of fluorescent aptamer sensor and application of fluorescent aptamer sensor in novel coronavirus detection
  • Construction method of fluorescent aptamer sensor and application of fluorescent aptamer sensor in novel coronavirus detection
  • Construction method of fluorescent aptamer sensor and application of fluorescent aptamer sensor in novel coronavirus detection

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] In the enzyme-free catalytic system based on the conformational entropy change to drive the forward reaction, figure 1 toehold shown in with The length will affect the initiation of the strand displacement reaction.

[0066] This embodiment uses three areas with The number of bases combined with different S1 strands, the difference between the three is:

[0067] In the first S1 chain: 6nt / 4nt.

[0068] In the second S1 strand: 5nt / 5nt.

[0069] In the third S1 strand: 4nt / 6nt.

[0070] The three S1 strands were pre-reacted with the OP strand and the SB strand in a 1:1:1 mixed system at 35-45°C for 30-45 min to form three sets of different substrate probes (S) with a stable triple-strand structure. The sets are substrate probes S①, S②, and S③ respectively. The selection of embodiment 2 triple-strand composite substrate probes

Embodiment 2 3

[0070] The three S1 strands were pre-reacted with the OP strand and the SB strand in a 1:1:1 mixed system at 35-45°C for 30-45 min to form three sets of different substrate probes (S) with a stable triple-strand structure. The sets are substrate probes S①, S②, and S③ respectively. The selection of embodiment 2 triple-strand composite substrate probes

[0071] Three sets of substrate probes (200nmol / L) prepared in Example 1 were reacted with C chain (200nmol / L) and F chain (200nmol / L) respectively to generate W chain, and the blank control group test was carried out simultaneously.

[0072] The generated W chain electrophoresis, the result is as follows figure 2 , figure 2 It is the product electrophoresis of the reaction system of different S probes, in which channels 1-3: S①, channels 4-6: S②, channels 7-9: S③, and channels 1, 4, and 7 are blank control groups.

[0073] The results showed that when the triple-stranded complex substrate probe toehold and domain When t...

Embodiment 3

[0075] The selection of embodiment 3F chain concentration

[0076] The F chain acts as a dye chain to provide reaction power, and its suitable concentration will affect the specificity and reaction efficiency of the chain displacement reaction driven by conformational entropy.

[0077] Get the three-chain composite substrate probe S of embodiment 2 (200nmol / L), C chain (200nmol / L) and the F chain of different concentrations, the concentration of F chain (100nmol / L, 200nmol / L, 300nmol / L, 400nmol / L).

[0078] S② (200nmol / L), C chain (200nmol / L) and the different concentrations of F chain were reacted to generate W chain, and the blank control test was carried out at the same time.

[0079] The generated W chain electrophoresis, the result is as follows image 3 , image 3 Electropherograms of products in reaction systems with different concentrations of F chain. Among them, channels 1~3: the concentration of F chain is 400nmol / L, channels 4~6: the concentration of F chain is...

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Abstract

The invention discloses a preparation method of a fluorescent aptamer sensor based on conformation entropy driving DNA assembly reaction. The preparation method comprises the following steps: (1) preparing g-CNQDs; (2) preparing Zn-MOF; (3) preparing a g-CNQDs@Zn-MOF solution: synthesizing the g-CNQDs@Zn-MOF solution from the Zn-MOF and the g-CNQDs; (4) carrying out biological functionalization on the g-CNQDs@Zn-MOF: connecting an MB chain to the g-CNQDs@Zn-MOF; and (5) forming the fluorescent aptamer sensor. The sensor is used for detecting the novel coronavirus, and analysis can be completed within 30 min.

Description

technical field [0001] The application belongs to the field of biomedicine, and relates to a construction method of a fluorescent aptasensor and its application in the detection of novel coronavirus. Background technique [0002] The coronavirus research group of the International Split Standard Virus Committee named the coronavirus that causes COVID-19 pneumonia as severe acute respiratory syndrome coronavirus 2 (Severe Acute Respiratory Syndrome Coronavirus 2, SARS-CoV-2), which is mainly transmitted by droplets and close contact . As the virus continues to mutate, the infectious ability of SARS-CoV-2 has increased. [0003] At present, the standard method for detecting SARS-CoV-2 in food and related places is the real-time fluorescent RT-PCR method, which is suitable for professionals to operate under laboratory conditions, which takes a long time and cannot meet the needs of rapid on-site analysis. It is difficult to timely detect SARS-CoV-2 in contaminated food and re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/533G01N33/569G01N21/64
CPCG01N33/533G01N33/56983G01N21/64G01N2333/165G01N2469/10Y02A50/30
Inventor 周琛邹海民林驰量昝昊成徐夏彦睿胡余瑶
Owner SICHUAN UNIV
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