Method for rapidly detecting vibrio mimicus in clinical blood sample

Abstract

The invention discloses a method for rapidly detecting vibrio mimicus in a clinical blood sample. The method comprises the following steps: S1, acquiring a blood sample to be detected, and extracting a DNA template; and S2, performing real-time fluorescence RPA detection by utilizing a combination of a primer and a probe, and judging whether the biological sample is infected with the vibrio mimicus or not according to an amplification result, wherein the primer and the probe are designed by taking a related virulence factor vmhA gene in vibrio mimicus as a target spot. The method has extremely high specificity and sensitivity, and a positive result can be obtained within the shortest 3.32 min by a real-time fluorescence RPA method at a relatively high concentration (3.5*10 < 6 > cfu / mL). The real-time fluorescence RPA method has very good tolerance to inhibitory components in a plasma sample, and can be used for directly amplifying a clinical plasma sample treated by a simple boiling method, so that the whole detection time is shortened. Besides, the real-time RPA method can be combined with portable equipment, and is a very useful method for monitoring mimicry vibrio infection in clinical diagnosis of a mimicry vibrio outbreak site.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for rapidly detecting Vibrio mimicus in clinical blood samples. Background technique [0002] Vibrio mimicus, as one of the main pathogenic bacteria in the genus Vibrio, is widely distributed in natural environments including freshwater and seawater, as well as in wild or farmed aquatic animals in contact with it, often causing human and aquatic animal illness. ill. Because of its morphology, physiological and biochemical characteristics, clinical manifestations of disease, and genome characteristics are very similar to Vibriocholera, it was considered to be the same pathogen in the early days. In the process of rapid expansion and reproduction, Vibrio mimeticum can produce a variety of virulence factors including hemolysin, enterotoxin, and metalloproteinase. Therefore, people often cause bacterial enteritis after eating aquatic products infected by Vibrio mimicus, leadin...

AI Technical Summary

Problems solved by technology

However, due to the inherent shortcomings of PCR technology itself, such as the need for a complete set of equipment for verification and amplification detection, certain experimental conditions and complex operating procedures, etc., it greatly limits its actual diagnostic detection in Vibrio mimeticus (especially rapid Application in grass-roots on-site diagnostic testing)

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