Detection primer group, detection kit, and detection method used for simultaneous detection of three pathogenic bacteria of marine fishes

Abstract

The invention discloses a detection primer group, a detection kit, and a detection method of nocardia seriolae, vibrio harveyi, and streptococcus iniae. The detection primer group comprises a primer pair Ns-mce, a primer pair Vh-tox, and a primer pair Si-its; the nucleotide sequences of the primer pairs are represented by SEQ ID NO.1-6. The detection kit comprises above detection primer group. According to the detection method, the detection kit is used for detection; samples are subjected to pretreatment, genome DNA is extracted as sample group DNA, and PCR is adopted to detect the samples. A triplex PCR detection method is established for tracking measurement of nocardia seriolae, vibrio harveyi, and streptococcus iniae at different periods, avoiding transmission of pathogenic bacteria, and improving rapid inspection and quarantine of pathogenic bacteria in aquatic products; and the invention belongs to the field of aquatic cultured animal pathogenic bacteria detection.

Description

technical field [0001] The invention belongs to the field of detection of pathogenic bacteria in aquaculture animals, and in particular relates to a detection primer set, a detection kit and a detection method for Nocardia spp., Vibrio harveii and Streptococcus iniae. Background technique [0002] Nocardiaseriolae, Vibrioharveyi, and Streptococcusiniae are three common pathogenic bacteria in mariculture fish, of which Nocardiaseriae and Streptococcus iniae belong to Gram positive bacteria, and Vibrio harveii is a Gram-negative bacteria. These three bacteria are widely distributed in the marine environment and aquaculture environment, and can be transmitted through bait, water environment, etc. They have certain differences in biological characteristics and pathogenicity, but they can all cause a large number of deaths of a variety of aquaculture animals , causing severe economic losses to the aquaculture industry. [0003] Yellowtail Nocardia can grow at 25-40°C, the optim...

AI Technical Summary

Problems solved by technology

[0006] Based on the existing technology, the detection of pathogenic bacteria in aquatic animals mostly adopts methods such as isolation, cultivation and identification of disease-like bacteria. The operation process is complicated, time-consuming, and the detection sensitivity is low. The result is uncertain and often delays the diagnosis of the disease.

In response to sudden disease events, it cannot meet the requirements of timely diagnosis, high sensitivity and specificity, accurate test results, and a large number of samples.

For the rapid detection of Nocardia spp., Vibrio harveii and Streptococcus iniae, domestic and foreign scholars have established bacterial culture identification and PCR detection methods for single pathogenic bacteria. However, in the process of seawater aquatic animal breeding, the In the prevention and control of related diseases, the sample size is large and the time is tight, and it is necessary to quickly detect multiple pathogenic bacteria at the same time, but a single pathogenic bacteria PCR detection method or a general bacterial isolation method cannot meet the above requirements

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