Multiplex fluorescent PCR kit for detection of vibrios in water body and detection method.

Abstract

The invention discloses a quadruple fluorescent PCR kit for detection of pathogenic vibrios in a water body and a detection method. According to the method, innovative bacterium enrichment reagent technology, DNA extraction reagent technology, specific fluorescent probe hybridization PCR detection technology and multiplex PCR technology are employed, bacterium enrichment culture of a sample is not needed, false positive interference is minimized as much as possible on the premise that high sensitivity is maintained, and four kinds of vibrios can be detected at one time. The invention mainly has the following beneficial effects: no need for bacterium enrichment of the sample in advance; high sensitivity and good specificity in detection of bacteria; reduction of the false positive incidence in conventional PCR amplification; and realization of rapid, accurate and specific detection of vibrio vulnificus, vibrio alginolyticus, vibrio mimicus and vibrio flurialis at one time.

Description

technical field [0001] The invention relates to the detection of Vibrio, in particular to a multiple fluorescent PCR kit and detection method for detecting Vibrio in water. Background technique [0002] At present, the traditional detection method of pathogenic Vibrio is still isolated and identified through microbial culture, which is not perfect enough. It needs to go through multiple screening steps, requires a variety of media, and takes a long time; there is also a monoclonal antibody using Vibrio The detection method requires picking a single bacterial colony and then using immunological methods to identify it. The specificity is high, but the time required for the detection of the two methods is relatively long. In terms of molecular biology testing, the FDA standard is still to design a PCR program for the purpose of detecting a pathogenic Vibrio. Domestic detection reagents design PCR detection method can only detect 3 kinds of Vibrio at the same time, and the focu...

AI Technical Summary

Problems solved by technology

[0002] At present, the traditional detection method of pathogenic Vibrio is still isolated and identified through microbial culture, which is not perfect enough. It needs to go through multiple screening steps, requires a variety of media, and takes a long time; there is also a monoclonal antibody using Vibrio The detection method requires picking a single colony and then using immunological methods to identify it after a large number of bacteria. The specificity is high, but the time required for the detection of the two methods is relatively long