Vibrio quantity detecting kit, preparation method thereof and vibrio quantity detecting method

A kit and vibrio technology, applied in the direction of biochemical equipment and methods, microbiological determination/inspection, etc., can solve the problem of difficult to meet the urgent needs of production and long-term use of large-scale breeding, only qualitative analysis, medium storage time To prevent and control vibriosis, facilitate disease prevention, and facilitate production management

Inactive Publication Date: 2014-04-23
ZHUHAI QIANGLONG BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the conventional method for detecting the number of Vibrio is the plate counting method, which consumes 2-3 days each time, and the medium storage time is short, which is difficult to meet the urgent needs of production and long-term use of large-scale cultivation; DAN probe technology requires high technical requirements. And only qualitative analysis can be performed; fluorescent quantitative PCR technology has good specificity and high sensitivity, but it can only be used for laboratory testing and water body testing

Method used

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  • Vibrio quantity detecting kit, preparation method thereof and vibrio quantity detecting method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] In this embodiment, a kit for detecting the number of Vibrio is provided, which includes Vibrio diluent and solid medium.

[0030] The vibrio dilution is a sodium chloride solution of 0.009g / ml, and the pH value of the sodium chloride solution is 8.5-8.7;

[0031] The formula of the solid medium is: yeast extract 3g, peptone 6g, sucrose 12g, sodium thiosulfate 6g, sodium citrate 6g, sodium taurocholate 1.8g, ox gall powder 3g, sodium chloride 6g, ferric citrate 0.6g, 2% bromothymol blue solution 12ml, 1% thymol blue solution 2.4ml, agar 9g.

[0032] The present embodiment also provides a preparation method of a kit for Vibrio quantity detection, comprising the following steps:

[0033] 1. Preparation of centrifuge tubes containing Vibrio dilution:

[0034] (1) Weigh 9g of sodium chloride and pour it into a beaker filled with 800ml of distilled water, stir evenly, adjust the pH value of the solution in the beaker to 8.5-8.7 with hydrochloric acid or sodium hydroxide, a...

Embodiment 2

[0056] In this embodiment, a kit for the detection of the number of Vibrio is provided, which is different from the kit for the detection of the number of Vibrio provided in Example 1 in that:

[0057] The formula of the solid medium is: yeast extract 5g, peptone 10g, sucrose 20g, sodium thiosulfate 10g, sodium citrate 10g, sodium taurocholate 3g, ox gall powder 5g, sodium chloride 10g, ferric citrate 1g , 2% bromothymol blue solution 20ml, 1% thymol blue solution 4ml, agar 15g;

[0058] The present embodiment also provides a preparation method of a test kit for the detection of the number of Vibrio, compared with the preparation method of the test kit for the detection of the number of Vibrio provided in the embodiment, the difference is that:

[0059] Preparation of Petri dishes with solid medium:

[0060] (1) Take 5g of yeast extract, 10g of peptone, 20g of sucrose, 10g of sodium thiosulfate, 10g of sodium citrate, 3g of sodium taurocholate, 5g of ox gall powder, 10g of so...

Embodiment 3

[0063] Effect experiment of the solid medium provided by the invention:

[0064] The known bacterial liquid 10 3 After the order of cfu / ml, take 100 μl, respectively spread on three solid medium inverted plates of S1 (solid medium used in the present invention), S2 (common nutrient agar) and S3 (TCBS medium), and incubate at 25°C for 18 h , to count the number of colonies formed.

[0065] Colony formation rate = actual number of clones / theoretical number of cells before coating * 100%

[0066] The solid medium culture effect is shown in Table 1:

[0067] Media name Theoretical number of bacteria before coating The actual number of clones formed Colony formation rate (%) S1 120 115 95.8 S2 120 113 94.2 S3 120 112 93.3

[0068] Conclusion: The cloning rate of the medium S1 provided by the invention as a solid medium is greater than 95%, and the detection of the number of Vibrio contained in the water body or tissue is more accurate. ...

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Abstract

The invention provides a vibrio quantity detecting kit comprising a vibrio diluent and a solid culture medium, wherein the vibrio diluent is a 0.009g / ml sodium chloride solution; and the solid culture medium comprises 3-5g of yeast cream, 6-10g of peptone, 12-20g of sucrose, 6-10g of sodium thiosulfate, 6-10g of sodium citrate, 1.8-3g of glycocholate sodium, 3-5g of ox gallbladder powder, 6-10g of sodium chloride, 0.6-1g of ferric citrate, 12-20ml of 2% bromothymol blue solution, 2.4-4ml of 1% thymol blue solution and 9-15g of agar according to a formula. The invention also provides a preparation method of the vibrio quantity detecting kit and a vibrio quantity detecting method realized by using the vibrio quantity detecting kit. The vibrio quantity detecting kit, the preparation method thereof and the vibrio quantity detecting method provided by the invention have the following technical effects of simplicity in operation, shorter detecting time and capability of effectively preventing and controlling vibriosis and providing a certain basis and guidance for clinical medication.

Description

technical field [0001] The invention relates to the technical field of aquaculture, in particular to a detection kit for the number of vibrio and its preparation method and detection method, more specifically to a vibrio detection kit for detecting aquatic products or water samples in freshwater or seawater cultivation A kit for quantitative detection, a preparation method and a detection method thereof. Background technique [0002] In the prior art, with the rapid development of the aquaculture industry in recent years, the problem of aquaculture animal diseases has become increasingly prominent, among which bacterial diseases have caused huge losses to the fishery economy worldwide. Vibrio is a large group of Gram-negative bacteria with short, terminal flagella, motile, non-spore-shaped, short rods curved into arcs, belonging to the Vibrio family and the genus Vibrio. A large number of studies have shown that a variety of Vibrio can cause epidemics and explosive deaths o...

Claims

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Application Information

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IPC IPC(8): C12Q1/04
Inventor 吴月喜陈玉婷陈小星李春震陈明泽杨先乐梁成锦吴汝泳吴明冯学峥陈桓梁丽丹孙玉平
Owner ZHUHAI QIANGLONG BIO TECH
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