A group of oligonucleotide aptamers that specifically recognize Bacillus cereus

Abstract

The invention discloses an oligonucleotide aptamer set (Sp15, Sp16) for specifically identifying Bacillus cereus, belonging to the fields of food sanitation and clinical medicine detection. In order to overcome the defect of no Bacillus cereus nucleotide aptamer in the prior art, a competitive Cell-SELEX technique is utilized to perform screening, amplification, enzyme digestion, sequencing and affinity and specificity analysis on the in-vitro synthetic random single-chain oligonucleotide library by using a complete active Bacillus cereus cell as the target, thereby obtaining the oligonucleotide aptamer set for specifically identifying Bacillus cereus. The screened aptamers have the characteristics of high affinity, high specificity, stable properties, low cost and the like, are convenient for in-vitro synthesis, can be used for detecting Bacillus cereus in the food and environment by marking the functional group or fluorescent dye, and provide a new option for the antibody-dependent detection method at present.

Description

technical field [0001] The invention relates to the field of food safety biotechnology, in particular to screening an oligonucleotide with high specificity and high affinity for Bacillus cereus using the SELEX technology (exponentially enriched ligand system evolution technology) in molecular biology technology The acid aptamer provides scientific basis and theoretical basis for the application of the nucleic acid aptamer in detecting Bacillus cereus. Background technique [0002] Bacillus cereus is a food-borne pathogen widely distributed in nature. It often exists in soil, dust and sewage, plants and various raw and cooked foods containing more starch. It has been removed from including The bacterium has been isolated from a variety of foods including meat, dairy products, vegetables, fish, potatoes, soy sauce, pudding, fried rice, and various desserts. The report of Bacillus cereus as a food-borne pathogenic bacterium is relatively rare. There are many, and the detection r...

AI Technical Summary

Problems solved by technology

Traditional bacterial culture and biochemical identification are cumbersome and time-consuming; molecular biology detection methods such as PCR methods are fast and accurate, but their influencing factors are complex and easily affected by operating conditions, with poor repeatability and specificity cannot be guaranteed; Chemical detection has the characteristics of strong specificity, accuracy and sensitivity, but the process of preparing specific antibodies is complicated and time-consuming, and the prepared antibodies have poor stability and are easily affected by environmental factors such as temperature

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