Nucleic acid aptamer for detecting clenbuterol hydrochloride and screening method and application thereof

Abstract

The invention provides a nucleic acid aptamer for detecting clenbuterol hydrochloride. The nucleic acid aptamer is selected from one of following nucleotide sequences shown in SEQ ID NO.1; the nucleicacid aptamers with same lengths are cut from both ends of the nucleotide sequence shown in SEQ ID NO.1, and the cutting length is 1 to 11bp; the nucleic acid aptamers with same cutting lengths of 1 to 11bp at the two ends respectively have the corresponding nucleotide sequences shown in SEQ ID NO.12 to SEQ ID NO.2. The invention also provides a screening method and application of the nucleic acidaptamer. The nucleic acid aptamer has the advantages that the specificity is strong, and the nucleic acid aptamer does not generate crossing reaction with clenbuterol hydrochloride analogues; the affinity is high, and the affinity constant is (42.17+ / -8.97) nM; the sensitivity is high; the minimum detection limit of clenbuterol hydrochloride is 1.2ng / mL.

Description

technical field [0001] The invention relates to the field of biomedical technology, in particular to a nucleic acid aptamer for detecting clenbuterol hydrochloride and its screening method and application. Background technique [0002] Clenbuterol hydrochloride (Clenbμterol, CBL), is a β-adrenoceptor agonist, originally used for the treatment of asthma. After CBL enters the livestock and poultry body, it can obviously promote the growth of animals and increase the lean meat rate. Human consumption of the edible parts of animals containing clenbuterol can cause food poisoning. The Ministry of Agriculture of my country strictly prohibited the application of adrenaline in animal production in March 1997 (Nongmufa [1997] No. 3 document). However, driven by economic interests, incidents of CBL exceeding the standard in livestock and poultry products occur from time to time. Therefore, it is necessary to monitor the residues of CBL in livestock and poultry products for a long t...

AI Technical Summary

Problems solved by technology

The bottleneck problem lies in: First, compared with biological macromolecules, it is relatively difficult to screen and identify nucleic acid aptamers for small molecule hazards

First, the number of screening rounds is twice that of this patent, which takes too long

Second, it uses the method of TA cloning and sequencing, and the amount of sequence information is relatively small

Third, it uses the fluorescent aptamer-graphene oxide method for application, the aptamer needs fluorescent modification, and the detection cost is high

Images