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Nucleic acid aptamers for specifically recognizing Staphylococcus aureus enterotoxin A

A staphylococcal intestinal and nucleic acid aptamer technology, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems of difficult to obtain antibodies, expensive preparation costs, unutilized, etc., and achieve operational The effect of convenience, low cost and simple preparation method

Active Publication Date: 2013-07-31
JIANGNAN UNIV
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Problems solved by technology

[0004] At present, domestic and foreign reports on protein target nucleic acid aptamers screened by SELEX technology mainly include thrombin, IgE, HIV-1 reverse transcriptase, neuropeptide Y, ricin, botulinum toxin, prostate Specific antigen, DNA-dealkylation enzyme and Staphylococcus aureus enterotoxin B, etc., there is no report on the nucleic acid aptamer sequence of Staphylococcus aureus enterotoxin A
However, SEA antibodies are difficult to obtain and expensive to prepare, and are subject to many conditions when used. Compared with antibodies, aptamers have more significant advantages, but their advantages have not been utilized in research in this area, so related research work needs to be strengthened urgently

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  • Nucleic acid aptamers for specifically recognizing Staphylococcus aureus enterotoxin A
  • Nucleic acid aptamers for specifically recognizing Staphylococcus aureus enterotoxin A
  • Nucleic acid aptamers for specifically recognizing Staphylococcus aureus enterotoxin A

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Embodiment Construction

[0013] The inventors of this case have found through long-term and practice that the interaction between single-stranded nucleic acid bases can make it form many possible spatial structures, including hairpins, pseudoknots, convex loops, and G-quadruplexes. Various target molecules undergo conformational binding. Based on this conformational recognition, a sufficient capacity (10 14 -10 15 ) nucleic acid library can contain as much three-dimensional structure as possible, so it is possible to screen high-affinity aptamers of any kind of target substance therefrom. Using SELEX technology, the target toxin SEA has an exponential evolutionary enrichment effect on each round of screening nucleic acid library. After dozens of rounds of repeated screening, reverse screening and negative screening are combined, and finally a nucleic acid aptamer sequence with high affinity and good specificity is obtained. .

[0014] Based on the above findings, the inventors of the present case p...

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Abstract

The invention discloses nucleic acid aptamers for specifically recognizing Staphylococcus aureus enterotoxin A, belonging to the technical field of biology of food safety. The invention aims to overcome the defect of no nucleic acid aptamers for Staphylococcus aureus enterotoxin A in the prior art. After 12 times of repeated analysis and verification of screening as well as affinity and specificity testing, the nucleic acid aptamers with favorable affinity and specificity for Staphylococcus aureus enterotoxin A are obtained. The invention provides specific and efficient recognition aptamers for analyzing and detecting the Staphylococcus aureus enterotoxin A in food or environment, and provides a new option for developing and substituting the existing method for detecting Staphylococcus aureus enterotoxin A on the basis of antibodies.

Description

technical field [0001] The invention relates to the field of food safety biotechnology, specifically refers to the preparation of a group of highly specific and high Affinity nucleic acid aptamers provide scientific basis and theoretical basis for the development of new detection technology based on nucleic acid aptamer recognition-Staphylococcus aureus enterotoxin A. Background technique [0002] Staphylococcus aureus Enterotoxins (SEs for short) are a group of small molecular weight (26-30kDa) heat-resistant protein toxins secreted by Staphylococcus aureus into the extracellular space. Food poisoning caused by this enterotoxin accounts for a very high proportion of bacterial food poisoning and is a worldwide public health problem, of which more than 75% are caused by Staphylococcus aureus enterotoxin A (SEA), followed by SED, SEC, and SEB. The poisoning symptoms of this type of enterotoxin are mainly manifested as nausea, vomiting, diarrhea and other adverse reactions wi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/115C12Q1/68C07K14/31
Inventor 王周平黄玉坤夏雨陈秀娟
Owner JIANGNAN UNIV
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