RPA reaction system suitable for rapid detection of Photobacterium mermaida mermaid subspecies

A technology of photobacteria and reaction system, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of no obvious geographical or host specificity, achieve good interspecies recognition, reduce condition requirements, and conserve sex high effect

Active Publication Date: 2021-08-10
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

[0003] In the previous research process, we have found that the mermaid subspecies of Photobacterium mermaid is ubiquitous in the marine aquaculture environment of our country, and can infect many kinds of fish such as grouper, flounder flounder, and flatfish Xu's. There is no obvious geographical area or host-specific

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  • RPA reaction system suitable for rapid detection of Photobacterium mermaida mermaid subspecies
  • RPA reaction system suitable for rapid detection of Photobacterium mermaida mermaid subspecies
  • RPA reaction system suitable for rapid detection of Photobacterium mermaida mermaid subspecies

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Embodiment 1

[0033] From the pathogen bank of aquatic animals in this laboratory, 10 strains of different pathogenic bacteria (Table 1) were randomly selected and detected by the RPA reaction system of the present invention. Take out the preservation solution of 10 strains of pathogenic bacteria from the -80°C ultra-low temperature refrigerator, put it at 4°C, let the bacteria liquid melt naturally, draw 30 μL and streak on the TSB solid medium, and incubate at 28°C for 24-36 hours. After colony formation, pick a single colony and streak again on a new TSB solid medium for purification at 28°C. After purification and cultivation for 3 times, a single colony was scraped and suspended in 0.5 mL of sterile 1.5% NaCl solution to prepare a bacterial suspension.

[0034] Table 1 The reference strains used for the specific detection of Photobacterium mermaida mermaid subspecies

[0035]

[0036] Add 100 μL of 2×LysisBuffer tissue lysate from Shanghai Chuangkun Biotechnology Co., Ltd. to the pre...

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Abstract

The invention belongs to the technical field of mariculture disease prevention and control, and in particular relates to an RPA reaction system suitable for rapid detection of Photobacterium mermaid subspecies mermaid. Specific primers were designed to carry out the amplification reaction of recombinase polymerase, and after gel electrophoresis verification of the product, it was judged qualitatively whether the detection sample contained Photobacterium mermaida subsp. The invention provides a stable, specific and highly efficient Recombinase Polymerase Amplification (RPA) reaction system of Photobacterium mermaidi subsp. The research on RPA detection technology of Photobacterium mermaid subspecies has laid a technical foundation for the accurate and rapid detection of RPA of Photobacterium mermaid subspecies.

Description

technical field [0001] The invention belongs to the technical field of mariculture disease prevention and control, and in particular relates to an RPA reaction system suitable for rapid detection of Photobacterium mermaid subspecies mermaid. Background technique [0002] Photobacterium damselae (Photobacterium damselae) is widely distributed in the global marine environment, and it is an important pathogenic bacteria of Vibrioceae (Vibrionaceae) Photobacterium (Photobacterium), including P.damselae subsp.damselae , PDD) and P. damselae subsp. Infected people get sick. Photobacterium mermaid subspecies mermaid was first reported by American scientists in 1981, and then there were successive cases of this bacteria infecting different marine animals such as fish, crustaceans, molluscs, sea turtles, whales and dolphins around the world. In 1982, The Lancet magazine reported for the first time the case of human infection caused by Photobacterium mermaid subsp. mermaid. After th...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6844C12Q1/689C12Q1/04
CPCC12Q1/6844C12Q1/689C12Q2531/119C12Q2521/507C12Q2537/1376C12Q2565/125
Inventor 张正于永翔陈京王印庚廖梅杰荣小军张浩
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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