Nucleic acid aptamer capable of specifically recognizing shigella, screening method and application thereof

A nucleic acid aptamer, Shigella technology, applied in biochemical equipment and methods, microbial determination/inspection, resistance to vector-borne diseases, etc., can solve time-consuming and arduous problems, and achieve low preparation cost and accuracy Diagnostic, stabilizing effect of single-stranded oligonucleotides

Active Publication Date: 2011-04-13
JIANGNAN UNIV
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

In essence, most of the surface antigens contained in the same genus are the same, with only slight differences, and it is obviously a time-consuming and arduous task to find these differences and prepare corresponding specific antibodies

Method used

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  • Nucleic acid aptamer capable of specifically recognizing shigella, screening method and application thereof
  • Nucleic acid aptamer capable of specifically recognizing shigella, screening method and application thereof
  • Nucleic acid aptamer capable of specifically recognizing shigella, screening method and application thereof

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Embodiment Construction

[0016] The following is the preparation method of nucleic acid aptamers that specifically bind to Shigella by SELEX technology and its application in the rapid detection of Shigella.

[0017] 1. Synthesize random single-stranded DNA library and primers (synthesized by IDT)

[0018] Random single-stranded DNA (ssDNA) library: 5′-ATAGGAGTCACGACGACCAGAA-(N40)-TATGTGCGTCTACCTCTTGACTAAT-3′, a random ssDNA library with a length of 87 nt was constructed, with fixed primer sequences at both ends and a random sequence of 40 bases in the middle, The storage capacity is 10 14 Above; Primer I: 5'-ATAGGAGTCACGACGACCAGAA-3'; Primer II: 5'-ATTAGTCAAGAGGTAGACGCACATA-3' The random ssDNA library and the two primers were prepared with TE buffer into a 100 μmol / L stock solution and stored at -20°C for later use.

[0019] 2. PCR amplification of double-stranded DNA (dsDNA) and single-stranded DNA (ssDNA) libraries

[0020] Before each round of screening, the ssDNA library was amplified into a ds...

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Abstract

The invention discloses a nucleic acid aptamer capable of specifically recognizing shigella, a screening method and application thereof. In the technical scheme, the single-stranded DNA oligonucleotide aptamer capable of specifically recognizing shigella is obtained through the SELEX (Systematic Evolution of Ligands by Exponential enrich-m ent) technology, and the aptamer can be converted to a report aptamer for detecting shigella through marking a marker such as fluorescein, so that the invention has wide application prospects of quick and accurate detection of shigella in food.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the preparation of a nucleic acid aptamer with high specificity and high affinity for Shigella using SELEX technology (exponentially enriched ligand system evolution technology) in molecular biology technology, Provide scientific basis and theoretical basis for the application of the nucleic acid aptamer in the detection of Shigella. Background technique [0002] Shigella dysenteriae is a kind of Gram-negative bacteria and is the most common pathogen of human bacillary dysentery, also known as Shigella dysenteriae. Bacillary dysentery is the most common intestinal infectious disease, with the most patients in summer and autumn. The source of infection is mainly patients and carriers, through oral infection through food and drinking water contaminated with Shigella. Shigella can secrete strong endotoxins, causing fever, mental disorders, and even toxic shock, destroying intestinal m...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/115C12N15/10C12Q1/68C12Q1/10
CPCY02A50/30
Inventor 王周平丁晓莹
Owner JIANGNAN UNIV
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