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Reagent kit and detection method for quickly detecting shark vibrio

A kit and reagent technology, applied in the field of kits for rapid detection of Vibrio shark, can solve the problems of tediousness and time-consuming, and achieve the effects of high sensitivity, convenient detection and good specificity

Inactive Publication Date: 2009-05-13
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the techniques for identification of Vibrio include: 1. Classical physiological and biochemical identification methods, which are cumbersome, time-consuming, and require operators to have good professionalism; 2. Bacteria rapid automatic identification systems, which require expensive instruments and equipment and Skilled operators; 3. Nucleic acid-based detection technology, including DNA probe and PCR technology (polymerase chain reaction technology), has the characteristics of rapidity, simplicity, and sensitivity compared with the above-mentioned detection technologies, but there is no See patents for PCR technology and detection kits for the detection of Shark Vibrio

Method used

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  • Reagent kit and detection method for quickly detecting shark vibrio
  • Reagent kit and detection method for quickly detecting shark vibrio
  • Reagent kit and detection method for quickly detecting shark vibrio

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1 makes the kit for quick detection of Shark Vibrio by following formula:

[0033] (a) Reagent A (enrichment medium): contains tryptone, beef extract, yeast extract and agar powder in a weight ratio of 5:3:1:18.

[0034] (b) Polymerase chain reaction (25 μl system) reagents: including reaction premix reagent B and reagent C,

[0035] Reagent B contains 10x PCR buffer (with MgCl 2 15mM), specific primer pair VcS and VcA, dNTP (a mixture of four deoxynucleotides), bovine serum albumin and sterile double distilled water, the contents are 2.5μl, 0.5μl (10μM), 0.5μl (10μM) , 0.5 μl (10 μMeach), 0.5 μl (5 mg / ml) and 19.375 μl.

[0036] Reagent C: Taq enzyme, 0.125 μl (5 units / μl).

[0037] (c) Electrophoresis and chromogenic reagents, including reagent D, reagent E and reagent F.

[0038] Reagent D: 50 times TAE (electrophoresis buffer), containing trihydroxymethylaminomethane-acetic acid and ethylenediaminetetraacetic acid, the concentrations are 0.04M and 0....

Embodiment 2

[0046] Embodiment 2 makes the kit for quick detection of Vibrio sharkus by following formula:

[0047](a) Reagent A (enrichment medium): contains tryptone, beef extract, yeast extract and agar powder in a weight ratio of 5:3:1:18.

[0048] (b) Reagents for polymerase chain reaction (50 μl system): including reaction premix reagent B and reagent C.

[0049] Reagent B contains 10x PCR buffer (containing MgCl 2 15mM), specific primer pair VcS and VcA, dNTP (a mixture of four deoxynucleotides), bovine serum albumin and sterile double distilled water, the contents are 5.0μl, 1.0μl (10μM), 1.0μl (10μM) respectively , 1.0 μl (10 μMeach), 1.0 μl (5 mg / ml) and 38.75 μl.

[0050] Reagent C: Taq enzyme, 0.25 μl (5 units / μl).

[0051] (c) Electrophoresis and chromogenic reagents, including reagent D, reagent E and reagent F.

[0052] Reagent D: 50 times TAE (electrophoresis buffer), containing trihydroxymethylaminomethane-acetic acid and ethylenediaminetetraacetic acid, the concentra...

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Abstract

The invention discloses an agent box and detecting method of shark vibrio, which comprises the following parts: enriched medium as agent A, polyase chain reaction with preblended agent B and agent C, electrophoretic and color agent with agent D, agent E and agent F. The primer doesn't augument signal for detected sample without shark vibrio, which possesses excellent atopy to monitor pathogenic agent in the breeding water.

Description

technical field [0001] The invention relates to a kit for rapidly detecting Vibrio shark and a method for rapidly detecting Vibrio shark using the kit. Background technique [0002] Vibrio is the main pathogen of marine animals. It has been found that Vibrio shark is one of the pathogens that cause a large number of deaths in near river oysters. It can also cause diseases of marine fish. This bacteria also exists in natural seawater. Monitoring Vibrio sharks in farmed animals and seawater is an important measure to prevent and control the diseases it causes. At present, the techniques for identification of Vibrio include: 1. Classical physiological and biochemical identification methods, which are cumbersome, time-consuming, and require operators to have good professionalism; 2. Bacteria rapid automatic identification systems, which require expensive instruments and equipment and Skilled operators; 3. Nucleic acid-based detection technology, including DNA probe and PCR tech...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04C12Q1/68G01N27/447G01N21/33
Inventor 张其中张占会
Owner JINAN UNIVERSITY
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