RESUMO
A visualized, rapid, simple method was developed based on loop-mediated isothermal amplification (LAMP) to detect Vibrio splendidus in aquatic products and aquaculture waters. TakingVibrio splendidus as the research object, and the toxR gene of Vibrio splendidus as the target gene, the boiling method was determined as a fast method suitable for the genomic DNA extraction of Vibriosplendidus. The optimized and screened primers could specifically detect Vibrio splendidus, and the sensitivity of nucleic acid concentration detection could reach 10
RESUMO
Vibrio splendidus is an opportunistic pathogen in aquaculture. It can infect a variety of aquaculture animals and has caused huge losses to the aquaculture industry. In this study, a novel and efficient method for detecting V. splendidus was developed by combining the exonuclease Ⅲ amplification strategy with a nucleic acid test strip developed based on gold nanoparticles-labeled DNA probe. The results could be directly visualized by naked eyes, and this system overcame the difficulty in preparation of the monoclonal antibody used in conventional immunostrip. Upon optimization of experimental conditions, the detection limit of the strip was 5 ng/mL for the synthetic oligonucleotide DNA fragment and 10 ng/mL for the actual genomic DNA sample of V. splendidus. This test strip was more sensitive compared with the PCR method and was specific for the detection of V. splendidus. The rapid preparation of nucleic acid strip and the efficient detection of V. splendidus open a new way for the prevention and control of aquatic diseases.