Genetic heterogeneity amongVibrio alginolyticusstrains, and design of a PCR-based identification method usinggyrBgene sequence

Abstract

Vibrio alginolyticus, a pathogen among humans and marine animals, is ubiquitous in marine environments. The aims of this study were to analyze the relationships between genetic diversity and origins, and to develop new primers based on the gyrB sequence to identify V. alginolyticus isolated from various sources. To determine the genetic diversity of this bacterium, an arbitrarily primed polymerase chain reaction (AP-PCR) technique was performed on 36 strains of V. alginolyticus isolated from diarrhea patients and from diseased marine animals and environments in southern Thailand. The results showed distinct DNA fingerprints of all strains, indicating that they are genetically heterogeneous. For species-specific identification of V. alginolyticus, primers targeting the gyrB gene of V. alginolyticus were developed. Thirty reference Vibrio spp., 13 non-Vibrio spp., and 160 strains of V. alginolyticus isolated from various sources in southern Thailand were used to evaluate the specificity of these primers. Our results showed that the gyrB primers could specifically identify V. alginolyticus from all sample types. In addition, the detection limit of the PCR was at least 95 pg of DNA template. Therefore, we concluded that the newly designed gyrB primers are rapid, highly sensitive, and specific to identify V. alginolyticus isolated from various sources.

Rapid visual nucleic acid detection of Vibrio alginolyticus by recombinase polymerase amplification combined with CRISPR/Cas13a

“…identi cation of pathogens is a prerequisite and basis for the treatment of infections. There have been many reports using qPCR to detect V. alginolyticus (Kang et al 2016;Bunpa et al 2018). Although this method can enable the early diagnosis of V. alginolyticus infections, it requires a high-purity template and a relatively complex pretreatment process.…”

A T3SS Regulator Mutant HY9901 ΔaraC of Vibrio alginolyticus Decreased the Expression of HopPmaJ and Provided Protection to Danio rerio as a Live-Attenuated Vaccine

“…It is one of the main pathogens of fish, shrimp, shellfish, and other mariculture animals (Bunpa et al, 2020). In addition to V. alginolyticus representing a conditional pathogen of fish, it poses a serious threat to human public health and threatens the Chinese aquaculture industry, with reports of it causing human food poisoning, otitis media, septicemia, and other diseases (Li et al, 2009;Bunpa et al, 2018). Due to the large-scale outbreak of V. alginolyticus and the long-term overuse and abuse of a large number of antibiotics and other chemical drugs, drug resistance of Vibrio is increasing daily with major implications to the aquaculture industry (Shahimi et al, 2021).…”