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Detection plate for high-flux screening of zoonotic pathogenic bacterium phage and application of detection plate

A zoonotic and phage technology, applied in the field of microorganisms, can solve the problems of high cost, time-consuming, and laborious, and achieve the effect of high cost

Active Publication Date: 2020-01-07
INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The traditional spot method for screening phage detection products has disadvantages such as low throughput, high cost, time-consuming, laborious, and inability to accurately identify and count with the naked eye

Method used

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  • Detection plate for high-flux screening of zoonotic pathogenic bacterium phage and application of detection plate
  • Detection plate for high-flux screening of zoonotic pathogenic bacterium phage and application of detection plate
  • Detection plate for high-flux screening of zoonotic pathogenic bacterium phage and application of detection plate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Preparation of 96 dot array Salmonella phage detection deep well plate

[0038] 1. Materials and methods

[0039] ①Cultivation of Salmonella: Streak inoculation of Salmonella on LB agar medium, and incubate at 37°C for 12 hours. Pick a single colony, place it in a sterilized tube containing 1ml of LB liquid medium, and incubate at 37°C with shaking at 180r / min for 6h, until the liquid becomes turbid and the bacterial solution reaches the exponential phase.

[0040] ②Double-layer plate activation of phage: Take a sterile 5ml EP tube, add 100μl of the cultured Salmonella solution, and also add 100μl of the corresponding Salmonella phage that has been diluted to a suitable gradient, and then add the semi-solid LB that has been cooled to 45-55℃ in advance for culture Base (NaCl 1g / ml, tryptone 1g / ml, yeast extract 0.5g / ml, 0.6% agar powder, distilled water) 3ml, quickly inverted and mixed to prevent semi-solid coagulation, poured into the LB solid medium prepared...

Embodiment 2

[0047] Example 2 Experiment of Screening Pathogenic Bacteriophage Using High-throughput Screening Zoonotic Bacteria Phage Detection Panel

[0048] 1. Materials and steps

[0049] ① Activate the Salmonella to be tested: Streak culture of 140 strains of Salmonella in LB agar medium, pick a single colony in LB liquid medium, shake overnight, and cultivate into a bacterial suspension.

[0050] ②Preparation of double-layer detection plate for Salmonella: Autoclave 15ml of LB solid medium, pour it into the 96-dot matrix detection plate, and place it horizontally to solidify. LB semi-solid medium is heated and melted, then packed into 5ml EP tubes, after autoclaving, cool to 45-55°C, add 100μl of the Salmonella liquid to be tested, shake it quickly and pour it, and spread it on the LB solid plate to make a Salmonella double layer detection board.

[0051] ③ Spotting phages: Add 90 μl of normal saline to each well of the 96-dot array Salmonella phage detection plate prepared above, ...

Embodiment 3

[0074] Example 3 Constructing a high-throughput phage screening system

[0075] (1) Efficient sampling express collection system for pathogenic bacteria

[0076] Prepare a sampling tube with a sterile transportation semi-solid medium inside, and a small shovel connected inside the tube cover, which can scrape the bacterial sample on the disease material, and then insert the transport medium; print a QR code on the outside of the tube cover, and its main information is Varms management The information on sampling farms and slaughterhouses determined by personnel and sampling personnel has been stored, and the sampling tubes are sorted and packed in ziplock bags. All sampling tubes are wrapped in bags and pasted with QR codes. The information is the same as that of the sampling tubes, increasing the amount of sampling tasks. The sampler uses the mobile APP to scan the QR code of the package to confirm the task. After scanning the QR code of each sampling tube, he only needs to e...

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Abstract

The invention discloses a detection plate for high-flux screening of zoonotic pathogenic bacterium phage and application of the detection plate. According to the detection plate, a phage library of zoonotic pathogenic bacteria is formed by screened phage according to splitting effects of the phage upon different zoonotic pathogenic bacteria, and in addition, the detection plate for high-flux screening of zoonotic pathogenic bacterium phage is designed and prepared according to the phage library. The detection plate comprises a phage box A, an inoculation box B and a detection box C. By adopting the detection plate, a high-flux phage screening system can be established, and a set of screening system with complete standardized operation, high-flux detection, automatic recognition and conversion into intelligent combinations of statistic data, information uploading and cloud monitoring on drug resistance of veterinarians and a phage prevention and control (Varms) databank can be formed.

Description

technical field [0001] The invention relates to a phage detection plate for high-throughput screening of zoonotic pathogens and its application, belonging to the field of microorganisms. Background technique [0002] Bacterial infection and pollution are economic and health problems in our country and even in the world. The widespread use of antibiotics for nearly half a century has led to the prevalence of multiple resistance to zoonotic pathogens, such as Salmonella, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus, Clostridium perfringens, and Clostridium difficile. Due to its medicinal properties, bacteriophages have great application potential in the prevention and treatment of bacterial infection and bacterial contamination, and have become an urgent need for contemporary medical and animal product production. [0003] Bacteriophage is a specific virus of bacteria, with high specificity of species and even strains, while veterinary clinic...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34C12M1/26C12Q1/02C12R1/42C12R1/19C12R1/445C12R1/22C12R1/385C12R1/92
CPCC12Q1/02Y02A50/30
Inventor 刘玉庆
Owner INST OF ANIMAL SCI & VETERINARY MEDICINE SHANDONG ACADEMY OF AGRI SCI
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