A group of oligonucleotide aptamers for identifying clenbuterol hydrochloride, salbutamol and ractopamine with high specificity

Abstract

The invention provides a group of oligonucleotide aptamers Apt-1, Apt-2 and Apt-3 which are capable of simultaneously identifying clenbuterol hydrochloride and salbutamol, an oligonucleotide aptamer CLB-2 which is capable of identifying the clenbuterol hydrochloride with high specificity, an oligonucleotide aptamer SAL-5 which is capable of identifying the salbutamol with high specificity and two oligonucleotide aptamers RAC-5 and RAC-6 which are capable of identifying ractopamine with high specificity. Through an SELEX (Systematic Evolution of Ligands by Exponential Enrichment) technology based on Fe3O4 magnetic nanoparticle separation, a random oligonucleotide library is immobilized on avidin-enveloped magnetic nanoparticles by virtue of a complementary chain of a biotinylation marker, and the oligonucleotide aptamers, which are high in specific affinity, are finally obtained by conducting screening by 16 turns. The aptamers are broad in application prospect; and the aptamers, by virtue of marker function genes or fluorescent dyes, are applicable to detection of the clenbuterol hydrochloride, the salbutamol and the ractopamine in food; therefore, a new choice is provided for existing detection methods which depend on antibodies.

Description

technical field [0001] The invention relates to the field of food safety biotechnology, in particular to screening a group of oligonucleotide aptamers that recognize clenbuterol hydrochloride and a group of aptamers that recognize albuterol by using SELEX technology (exponential enrichment ligand system evolution technology). The oligonucleotide aptamer and a group of oligonucleotide aptamers recognizing ractopamine provide scientific and theoretical basis for the application of oligonucleotide aptamer-based detection of clenbuterol in food. Background technique [0002] Clenbuterol, as a synthetic β-adrenergic receptor activator, can reduce animal fat content, increase lean meat percentage, promote animal growth, and reduce feed consumption, so it is used in animal husbandry. However, clenbuterol metabolizes slowly in animals, and accumulates in the human body through meat products, causing poisoning symptoms such as dizziness, fatigue, heart palpitations, and numb limbs, w...

AI Technical Summary

Problems solved by technology

However, since clenbuterol hydrochloride, salbutamol, and ractopamine are all small molecular haptens, which are not immunogenic, they need to be combined with BSA and other macromolecular carrier proteins to prepare complete antigens before stimulating animals to secrete antibodies. Therefore, antibody preparation Not only is it time-consuming, tedious and expensive, but the prepared antibodies are also susceptible to environmental factors such as temperature

However, there is no research report on the preparation method of oligonucleotide aptamers for clenbuterol hydrochloride, albuterol, and ractopamine

Images