Three oligonucleotide sequences for identification and detection of vibrio alginolyticus as well as preparation method and application thereof

Abstract

The invention discloses three oligonucleotide sequences for identification and detection of vibrio alginolyticus as well as a preparation method and an application thereof and relates to identification and detection of vibrio alginolyticus. The oligonucleotide sequences comprise SEQ ID No. 1-3 (sequence identity number 1-3). The method comprises the following steps: synthesizing an ssDNA (single-stranded deoxyribonucleic acid) oligonucleotide library (5'-TCA GTC GCT TCG CCG TCT CCT TC----N35----GCA CAA GAG GGA GAC CCC AGA GGG-3') for screening; mixing the oligonucleotide library with the vibrio alginolyticus and then performing SELEX (systematic evolution of ligands by exponential enrichment) screening; performing cloning and sequencing on an aptamer-enhanced library after completing the SELEX screening; performing interception in different lengths on high-copy ssDNA emerged in the sequencing result so as to get a series of new sequences, and further constructing complementary sequences of the new sequences; and performing affinity and specificity verification on the obtained new sequences and the complementary sequences thereof so as to get corresponding aptamers.

Description

technical field [0001] The invention relates to the identification and detection of Vibrio alginolyticus, in particular to three oligonucleotide sequences that can be used for the identification and detection of Vibrio alginolyticus and its preparation method and application. Background technique [0002] In the aquaculture industry, about 10% of aquaculture animals die from infectious diseases every year, among which the diseases caused by Vibrio infection account for a considerable proportion. Although chemicals and antibiotics can control its infection, drug prevention and control often have adverse consequences, such as accumulation and residue in aquatic animals, easy to produce drug-resistant strains and easy pollution of the environment. Therefore, a rapid detection technology for pathogenic Vibrio is established to Early prevention and control of the occurrence and prevalence of diseases is still the main means at present. [0003] At present, the detection method o...

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Problems solved by technology

Although chemicals and antibiotics can control its infection, drug prevention and control often have adverse consequences, such as accumulation and residue in aquatic animals, easy to produce drug-resistant strains and easy pollution of the environment. Therefore, a rapid detection technology for pathogenic Vibrio is established to Early prevention and control of the occurrence and prevalence of diseases is still the main means at present

[0003] At present, the detection method of Vibrio is mainly based on the Bergey's Bacteria Identification Manual, through the detection of the physiological and biochemical characteristics of microorganisms. Since there are many items to be tested, this method has a large workload and cumbersome operation.

Although the molecular biology method of 16SRNA has high accuracy, it needs to extract 16SRNA of pathogenic bacteria. Realize rapid detection, but because the prepared antiserum is a polyclonal antibody, false positives or false negatives may occur in practical applications. Even if monoclonal antibodies are used, their application is limited due to high preparation technology requirements and long cycle

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