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Aptamer sequence for specifically recognizing ribavirin and application thereof

An aptamer and specific technology, applied in fluorescence/phosphorescence, instruments, biochemical equipment and methods, etc., can solve problems such as low affinity, cumbersome operation, and antibody instability

Active Publication Date: 2021-05-18
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The existing chromatographic method for detecting ribavirin has the problems of expensive equipment and cumbersome operation, while the immunological method has the problems of long cycle, low affinity, low specificity, unstable antibody and difficult preservation

Method used

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  • Aptamer sequence for specifically recognizing ribavirin and application thereof
  • Aptamer sequence for specifically recognizing ribavirin and application thereof
  • Aptamer sequence for specifically recognizing ribavirin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Screening of aptamers that specifically recognize ribavirin

[0033] 1. Fe 3 o 4 Preparation and Characterization of Nanomagnetic Beads

[0034] 1.1 Aminated Fe 3 o 4 Preparation and Characterization of Nanomagnetic Beads

[0035] One-step preparation using hot solvent: Weigh 1.0g ferric chloride hexahydrate, add 2.0g anhydrous sodium acetate into 30ml ethylene glycol, shake gently to mix, then add 6.5g of 1,6-hexamethylenediamine, at 50 Stir vigorously in a water bath at ℃ until a uniform wine-red transparent solution is formed, quickly transfer the resulting solution to a 100mL high-pressure reaction kettle with a Teflon liner, heat at 198℃ for 6h, take out the reaction kettle, and cool at room temperature . After the reaction kettle is cooled, remove the supernatant under the action of an external magnetic field, add 100mL of absolute ethanol to rinse and transfer the obtained lower black solid to a beaker, perform magnetic separation after ultrasonic...

Embodiment 2

[0071] Example 2 The method of using specific aptamers to detect ribavirin content in chicken samples

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Abstract

The invention discloses an aptamer sequence for specifically recognizing ribavirin and application thereof, and belongs to the technical field of food safety biology. According to the invention, ribavirin is taken as a target, aptamers are enriched through multiple rounds of screening processes by utilizing a Capture-SELEX technology, screening processes are monitored by applying real-time fluorescent quantitative PCR, and all sequence information of an aptamer library is intuitively and comprehensively obtained through high-throughput sequencing, so that an aptamer sequence with the highest enrichment degree is obtained through screening; affinity and specificity of the obtained nucleic acid sequence are represented by a colorimetric method and a fluorescence method, and an aptamer sequence with good affinity and specificity is obtained. The aptamer sequence can be used for detecting the content of ribavirin in bodies of livestock and poultry breeding animals, food and medicines used by livestock and poultry breeding animals, and human blood.

Description

technical field [0001] The invention relates to an aptamer sequence specifically recognizing ribavirin and application thereof, belonging to the field of food safety biotechnology. Background technique [0002] Ribavirin is a broad-spectrum antiviral drug, which can not only inhibit the replication of RNA viruses, but also has a strong inhibitory effect on DNA viruses, and is often used in the treatment of respiratory diseases. However, the use of veterinary drugs in the treatment of viral diseases in animals has great safety risks, and the sale and use of veterinary drugs is prohibited. [0003] The abuse of antiviral drugs will not only cause the decline of the quality and flavor of farmed animals, but also lead to drug residues and drug resistance. Abuse of ribavirin in poultry farming can lead to drug resistance of hepatitis C virus. There are about 40 million HCV carriers in my country, with an average infection rate of about 3.2%. More than 70% of those infected will...

Claims

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Application Information

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IPC IPC(8): C12N15/115C12N15/10G01N33/53G01N21/64
CPCC12N15/115C12N15/1048G01N33/94G01N33/53G01N21/6428C12N2310/16G01N2021/6432
Inventor 段诺宋明燕王周平吴世嘉
Owner JIANGNAN UNIV
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