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Method and kit for quickly identifying vibrio alginolyticus by violent vibrio alginolyticus bacteriophage

A technology of vibrio alginolyticus and bacteriophage, which is applied in biochemical equipment and methods, determination/inspection of microorganisms, etc., can solve the problems of inability to identify taxa, low accuracy, complicated operation, etc., and achieve accurate and rapid identification and low cost. , the effect of high efficiency

Inactive Publication Date: 2018-04-20
XIAMEN CANCO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Create new problems: There will be more and more taxa that cannot be identified only by phenotypic characteristics, but must be combined with genotype determination to identify
[0004] The commonly used rapid detection methods for Vibrio alginolyticus mainly include molecular biology methods and immunodiagnostic techniques; however, the above methods have the disadvantages of complicated operation, high cost, and low accuracy.

Method used

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  • Method and kit for quickly identifying vibrio alginolyticus by violent vibrio alginolyticus bacteriophage

Examples

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Embodiment 1

[0039] In this embodiment, the method for rapidly identifying Vibrio alginolyticus using potent alginolytic Vibrio phage comprises the following steps:

[0040] Step A: Isolation and purification of host bacteria: Vibrio alginolyticus was isolated from a large number of diseased shrimp intestines of Penaeus vannamei.

[0041] Step B: Expansion culture of host bacteria: add 5% NaCl to LB liquid medium, autoclave at 121° C. for 20 minutes, and cool to room temperature to obtain a culture solution. Under aseptic conditions, a single colony of Vibrio alginolyticus on the seed preservation plate was picked and inoculated into the culture medium, and cultured at 30° C. and 150 rpm for 16 hours to obtain the culture medium of the host bacteria.

[0042] Step C: Isolation of Vibrio alginolyticus phages: Vibrio alginolyticus phages were isolated from culture ponds of Penaeus vannamei.

[0043] Step D: Expansion of Vibrio alginolyticus phage: the isolated Vibrio alginolyticus phage is ...

Embodiment 2

[0047] In this embodiment, the method for rapidly identifying Vibrio alginolyticus using potent alginolytic Vibrio phage comprises the following steps:

[0048] Step A: Isolation and purification of host bacteria: Vibrio alginolyticus was isolated from a large number of diseased shrimp intestines of Penaeus vannamei.

[0049] Step B: expansion culture of host bacteria: add 2% NaCl to LB liquid medium, autoclave at 121° C. for 20 minutes, and cool to room temperature to obtain a culture solution. Add 10% of the volume of the culture solution to the culture solution under sterile conditions 6 The CFU / mL Vibrio alginolyticus bacteria solution was cultured at 32° C. and 200 rpm for 10 hours to obtain a host bacteria culture solution.

[0050] Step C: Isolation of Vibrio alginolyticus phages: Vibrio alginolyticus phages were isolated from culture ponds of Penaeus vannamei.

[0051] Step D: Expansion of Vibrio alginolyticus phage: the isolated Vibrio alginolyticus phage is expande...

Embodiment 3

[0055] In this embodiment, the method for rapidly identifying Vibrio alginolyticus using potent alginolytic Vibrio phage comprises the following steps:

[0056] Step A: Isolation and purification of host bacteria: Vibrio alginolyticus was isolated from a large number of diseased shrimp intestines of Penaeus vannamei.

[0057] Step B: Expansion culture of host bacteria: add 5% NaCl to LB liquid medium, autoclave at 121° C. for 20 minutes, and cool to room temperature to obtain a culture solution. Under aseptic conditions, a single colony of Vibrio alginolyticus on the seed preservation plate was picked and inoculated into the culture medium, and cultured at 28° C. and 120 rpm for 18 hours to obtain a culture medium of the host bacteria.

[0058] Step C: Isolation of Vibrio alginolyticus phages: Vibrio alginolyticus phages were isolated from culture ponds of Penaeus vannamei.

[0059] Step D: Expansion of Vibrio alginolyticus phage: the isolated Vibrio alginolyticus phage is ex...

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Abstract

The invention provides a method and a kit for quickly identifying vibrio alginolyticus by a violent vibrio alginolyticus bacteriophage, belonging to the technical field of strain identification of vibrio alginolyticus. The method comprises the following steps of 1) covering a strain to be detected on an LB agar plate, standing for 15 to 20min to obtain a plate to be detected; 2) dropwise adding the violent vibrio alginolyticus bacteriophage onto the plate to be detected to obtain an identification system; 3) performing stationary culture on the identification system obtained in the step 2) for8 to 16h at the temperature of 28 to 35 DEG C, if bacteriophage plaque appears, the strain to be detected is the vibrio alginolyticus. According to the method provided by the invention, the vibrio alginolyticus can be quickly and accurately identified; the method has the characteristics of high efficiency, low cost, time saving and labor saving; the accuracy rate reaches 100%; the method is applicable to preliminary screening and identification of a large number of vibrio alginolyticus.

Description

technical field [0001] The invention belongs to the technical field of identification of Vibrio alginolyticus strains, and in particular relates to a method and a kit for rapidly identifying Vibrio alginolyticus by using a potent Vibrio alginolyticus phage. Background technique [0002] Vibrio alginolyticus is a halophilic Gram-negative short bacillus without spores and capsules, belonging to the family Vibrio and the genus Vibrio. the first. It can cause wound infection, food poisoning, otitis media and other diseases to humans. At the same time, it is also an opportunistic pathogen for fish, shrimp, shellfish and other marine aquaculture animals. When the environment deteriorates, the body's immune function decreases, and alginolytic vibriosis is prone to outbreaks. [0003] Due to the complex and diverse physiological and biochemical characteristics and genetic characteristics of Vibrio alginolyticus, it is difficult to accurately identify it, and identification with tr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70
CPCC12Q1/70
Inventor 姜宗然付汉清郭立
Owner XIAMEN CANCO BIOTECH CO LTD
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