Rapid identification of related species of vibrio by gyrB gene degenerative primers
Dianfu Wang, Lidan Ma, Bei Xiong, Jinyan Zhu, Jing Lü, Ling-Li Meng, Yun‐Guo Liu
Abstract
Vibrio are ubiquitous in the marine environment, and some of them are pathogenic to marine organisms and humans. Due to the complexity and sensitivity of traditional phenotypic and biochemical tests, there is currently no method that could distinguish pathogenic Vibrio on the market. Therefore, this study developed new primers based on gyrB gene to identify Vibrio. We designed 7 pairs of PCR primers (DP1-DP7) and pre-selected them using 7 standard strains of Vibrio. The results showed that the efficiency of DP1 primers (VF1:5′-AARCARGGNCGTAACCGTAA-3′, VR1: 5′-HGGGTADCGRCGRCTCAT-3′) was better. After that, 15 standard strains of Vibrio, 4 non-Vibrio strains and 48 Vibrio strains were used to evaluate the specificity of DP1. Through agarose gel electrophoresis and sequencing analysis of the PCR amplified products. The results showed that the sequence consistency of DP1 primer amplification product was ≥99%, which was consistent with the target strain. This study proved that DP1 primer could be used as a universal primers for Vibrio, providing technical support for rapid screening and identification of Vibrio.