Aeromonas veronii rapid detection primer, kit and application

A technology of Aeromonas victorii and a detection kit, which is applied in the direction of microorganisms, methods based on microorganisms, and the measurement/inspection of microorganisms. It can solve the problems of time-consuming bacteria isolation and identification, economic losses for farmers, and complicated operations. To achieve the effects of programming and standardization, improving application reliability and operating specifications

Inactive Publication Date: 2015-04-22
天津市水产技术推广站
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Aeromonas verdeii is an opportunistic pathogenic bacterium that affects humans and animals. In recent years, freshwater cultured fish, such as catfish, carp, koi, Chinese soft-shelled turtle, and river crab, have been infected by Aeromonas verdeii. Reports have brought huge economic losses to the farmers
At present, for the detection of Aeromonas victorii in fish, traditional bacterial isolation and identification, serological reaction and PCR detection technology are mainly

Method used

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  • Aeromonas veronii rapid detection primer, kit and application
  • Aeromonas veronii rapid detection primer, kit and application
  • Aeromonas veronii rapid detection primer, kit and application

Examples

Experimental program
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Effect test

Embodiment 1

[0037] The rapid detection primer of Aeromonas victorii is composed of an outer primer and an inner primer, and the outer primer is composed of an outer primer upstream primer shown in SEQ ID NO.1 and an outer primer downstream primer shown in SEQ ID NO.2; the inner primer The primers consist of an inner primer upstream primer shown in SEQ ID NO.3 and an inner primer downstream primer shown in SEQ ID NO.4.

Embodiment 2

[0039] Rapid detection kit for Aeromonas verkirea, including:

[0040] (1) TE buffer solution, its composition is 20mmol / L Tris-HCl, 2mmol / L EDTA, solvent is distilled water, pH=8.0;

[0041] (2) Bst DNA polymerase: Bst DNA polymerase with a concentration of 8 U / μl;

[0042] (3) LAMP reaction solution, the composition of 24 μl LAMP reaction solution is: 2.5 μl 10×reaction buffer solution; 3 μl concentration is the dNTP of 2.5mmol / L; Primer upstream primer, 1 μl concentration of 5 μmol / L outer primer downstream primer shown in SEQ ID NO.2, 1 μl concentration of 40 μmol / L inner primer upstream primer shown in SEQ ID NO.3, 1 μl concentration of 40 μmol / L of the inner primer downstream primer shown in SEQ ID NO.4; 1 μl of the complex formed by calcein and manganese ions; 13.5 μl of DEPC water;

[0043](4) Positive control membrane, made by the following method: absorb the culture solution of Aeromonas vickerii to fully wet the FTA membrane, dry it at room temperature, put it in...

Embodiment 3

[0047] Rapid detection kit for Aeromonas verkirea, including:

[0048] (1)-(3) is the same as (1)-(3) of embodiment 2;

[0049] (4) Positive control membrane, made by the following method: absorb the culture solution of Aeromonas vickerii to fully wet the FTA membrane, dry it at room temperature, put it into a washing tube filled with 200 μl TE buffer and wash it twice with shaking , 3 minutes each time, fully dry;

[0050] (5) Negative control membrane, made by the following method: absorb DEPC water to fully wet the FTA membrane, then dry it at room temperature, put it into a washing tube filled with 200 μl TE buffer, shake and wash twice, each time for 3 minutes, and fully dry ;

[0051] (6) is the same as (6) of embodiment 2.

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Abstract

The invention discloses an aeromonas veronii rapid detection primer, a kit and application. The aeromonas veronii rapid detection primer is characterized by comprising an outer primer and an inner primer; the outer primer is composed of an outer primer upstream primer shown in SEQ ID NO.1 and an outer primer downstream primer shown in SEQ ID NO.2; the inner primer is composed of an inner primer upstream primer shown in SEQ ID NO.3 and an inner primer downstream primer shown in SEQ ID NO.4. According to the method, the period for detecting aeromonas veronii is short, the detection cost is low, sequencing and standardization of the detection process are achieved, operation is standard, and errors are not prone to being caused. The specificity is good. The dye built-in method is adopted, a reaction tube does not need to be opened after a reaction is finished, results can be observed directly with the naked eyes after the reaction tube is taken out, a follow-up to-be-detected sample is prevented from being polluted by amplified products, bacterial culture does not need to be carried out, and detection can be carried out only through fewer samples such as fish blood.

Description

technical field [0001] The invention relates to a rapid detection primer, kit and application of Aeromonas verkistan, belonging to the field of rapid detection of aquatic pathogens. Background technique [0002] Aeromonas veronii, also known as Aeromonas verona, Aeromonas vanlon, and Aeromonas veronii, is a genus of Gram-negative bacilli. It is a kind of Aeromonas discovered and identified in recent years. Aeromonas verdeii is an opportunistic pathogenic bacterium that affects humans and animals. In recent years, freshwater cultured fish, such as catfish, carp, koi, Chinese soft-shelled turtle, and river crab, have been infected by Aeromonas verdeii. Reports have brought huge economic losses to farmers. At present, for the detection of Aeromonas victorii in fish, traditional bacterial isolation and identification, serological reaction and PCR detection technology are mainly used. Bacterial isolation and identification is time-consuming and costly. Most of them are based on...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12R1/01
CPCC12Q1/6844C12Q1/689C12Q2531/119
Inventor 姚学良徐晓丽张振奎臧莉蔡琰郑艳坤任涵玮
Owner 天津市水产技术推广站
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