Four sets of oligonucleotide sequences for recognition and identification of vibrio anguillarum, and screening method of oligonucleotide sequences

Abstract

The invention discloses four sets of oligonucleotide sequences for recognition and identification of vibrio anguillarum, and a screening method of the oligonucleotide sequences, and relates to the technical field of recognition detection of vibrio anguillarum. Two ends of each oligonucleotide sequence are fixed sequences, intermediate sequences are arrayed according to the order of 5'-3' as follows: a first set contains two sequences of H1 and H2, a second set contains five sequences of H5, H6, H26, H38 and H33, a third set contains three sequences of H12, H25 and H42, and a fourth set contains a sequence of H28; and any of the oligonucleotide sequences in each set serves as the intermediate sequence of an aptamer to be used for vibrio anguillarum detection. The screening method of the sequences sequentially comprises the steps of random oligonucleotide library synthesis, combination, separation, PCR amplification and high-throughput sequencing which are circulated for 3-5 rounds, andfinally, the target sequences are obtained. The sequence length of the aptamer of the vibrio anguillarum is shortened, and the screening method is quick.

Description

technical field [0001] The invention relates to identification and detection technology of vibrio anguillarum, in particular to four groups of oligonucleotide sequences used for identification and identification of vibrio anguillarum and a screening method thereof. Background technique [0002] ( Vibrio anguillarum ) is a Gram-negative bacterium that is widely distributed and can infect more than 50 species of freshwater and marine fishes such as eels and salmonids and other aquatic animals. Vibrio eelium generally gathers in the rectal epithelium of the host, and then enters the host's lymphatic system and blood, causing aquaculture animals to infect diseases, such as intestinal inflammation, scale loss, abdominal swelling, systemic hemorrhage, etc., resulting in serious losses to the aquaculture industry. Therefore, accurate and rapid detection and identification of Vibrio anguillarum is very necessary, and it is also the premise and key to the treatment and prevention of...

AI Technical Summary

Problems solved by technology

In the first round of screening, there may even be too few aptamers that can bind, resulting in failure of PCR amplification, resulting in screening failure

[0005] In addition, in the existing Vibrio anguillarum aptamer screening, affinity determination is carried out in almost every round, the template of each round of PCR must be purified before PCR, and the PCR product of each round must be purified before it can be used as a screening library. For the next round of screening, these steps of affinity determination and purification greatly increase the workload of screening, increase the screening process and labor intensity, and greatly affect the efficiency of screening.

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