Rapid Identification of Vibriosis causative V. harveyi with Real-Time PCR
Project no. 22524 · Status: Kapanmış · 2021
Summary
Accurate and rapid characterization of the causative pathogen is of great importance in terms of diagnosis and treatment of the disease in order to prevent losses caused by microbial diseases in aquaculture. Studies have been carried out on the phenotypic characterization of Vibrio harveyi and the determination of the anatomical and histopathological findings that occur during the diseases it causes, by culturing many fish species and using traditional bacteriological techniques. With the widespread use of 16S rRNA analyzes in the recent past, there are many studies on the molecular diagnosis of this type of pathogen. However members of the Vibrionaceae family are a very difficult group to classify as genotypic and phenotypic. The aim of this study was to optimize an efficient method for the identification of V. harveyi based on the use of the toxR gene as a taxonomic marker. In this context, the primer and probe for the ToxR gene were designed specifically for V. harveyi. Real-Time PCR results, which took less than 4 hours from DNA isolation to amplification with the developed species-specific primer sets, revealed positive amplification of the ToxR gene fragment in V. harveyi isolates, while DNA from other Vibrio genus members did not amplify. The optimized method as a result of this project work enabled the rapid and precise differentiation and identification of V. harveyi and Vibrio sp. RT-PCR represents a significant advance in aquaculture by providing an easy, fast and cost-effective method for the identification of closely related pathogenic bacteria that are difficult to distinguish with traditional approaches.