Rapid detection kit for aeromonas schubertii and detection method

A detection kit, Aeromonas technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of low accuracy, time-consuming, etc., achieve moderate size, improve specificity, and reduce detection. effect of time

Active Publication Date: 2014-02-12
ZHEJIANG INST OF FRESH WATER FISHERIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the problems of time-consuming and low accuracy of the traditional identification method of A

Method used

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  • Rapid detection kit for aeromonas schubertii and detection method
  • Rapid detection kit for aeromonas schubertii and detection method

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0016] Embodiment one

[0017] Rapid detection kit for Aeromonas schubert, and use of the kit for specific detection of genomic DNA of different bacteria.

[0018] The kit consists of:

[0019] The composition of the DNA extraction reagent is 5mM Tris, 0.5mM EDTA pH 8.0 TE buffer; proteinase K, concentration 15mg / mL; 100% chloroform; 100% isopropanol; ethanol, concentration 70%; double distilled water ;

[0020] The PCR reaction reagents were divided into several reaction tubes, and each PCR reaction tube contained 2.5 μL of 10×PCR reaction buffer, 0.5 μL of dNTPs at a concentration of 10 μM, and 0.3 μL of Taq DNA polymerase at a concentration of 5 U / μL at a concentration of 10 μM 0.5 μL of specific oligonucleotide primer A, 0.5 μL of 10 μM specific oligonucleotide primer B, 19.2 μL of sterilized deionized water.

[0021] The detection method of the test kit provided by the present invention to Aeromonas schubert in the sample is to first use the DNA extraction reagent to c...

Example Embodiment

[0032] Embodiment 2

[0033] Aeromonas Schubert rapid detection kit, and the specific detection of genomic DNA of different bacteria using the kit.

[0034] The kit consists of:

[0035] The DNA extraction reagent consists of 15mM Tris, 1.5mM EDTA, pH 8.0 TE buffer; proteinase K, concentration 18mg / mL; 100% chloroform; 100% isopropanol; ethanol, concentration 70%; double distilled water ;

[0036] The PCR reaction reagents are distributed in several reaction tubes, each PCR reaction tube contains 2.5 μL of 10× PCR reaction buffer, 0.5 μL of dNTPs at a concentration of 10 μM, and 0.3 μL of Taq DNA polymerase at a concentration of 5 U / μL at a concentration of 10 μM. 0.5 μL of specific oligonucleotide primer A, 0.5 μL of 10 μM specific oligonucleotide primer B, and 19.2 μL of sterilized deionized water.

[0037] The method for detecting Aeromonas Schubert in a sample by the kit provided by the present invention is to first extract DNA from the sample with a DNA extraction reag...

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Abstract

The invention discloses a rapid detection kit for aeromonas schubertii. The rapid detection kit comprises a DNA (Deoxyribonucleic Acid) extraction reagent (1) and a PCR (Polymerase Chain Reaction) reagent (2), wherein the DNA extraction reagent comprises a TE (Tris-EDTA) buffer solution with 5-15mM of Tris and 0.5-1.5mM of EDTA (Ethylene Diamine Tetraacetic Acid), proteinase K with the concentration of 15-20mg/mL, 100% chloroform, 100% isopropanol, ethanol with the concentration of 70% and double distilled water; each PCR tube contains 2.5muL of 10*PCR buffer solution, 0.5muL of dNTPs with the concentration of 10muM, 0.3muL of Taq DNA polymerase with the concentration of 5U/muL, 0.5muL of specific oligonucleotide primers A with the concentration of 10muM, 0.5muL of specific oligonucleotide primers B with the concentration of 10muM, 19.2muL of sterilized deionized water, a primer A (5'-CAGCGAGGAGGAAAGGTTGGTGGT-3') and a primer B (5'-AAGCCACGCCTCAAGGGCACAA-3'). The rapid detection kit is used for detecting aeromonas schubertii.

Description

technical field [0001] The invention relates to a detection technology of target DNA fragments, in particular to a rapid detection kit for Aeromonas schubert and a method for detecting Aeromonas schubert with the kit. Background technique [0002] Aeromonas schubertii (Aeromonas schubertii) belongs to the Aeromonas family (Aeromonadaceae) Aeromonas (Aeromonas), is a Gram-negative bacillus, unipolar flagella, very active movement. The bacterium widely exists in fresh water, sea water, soil, fish and vertebrate intestines, and can cause infection after human contact. It is an important pathogenic bacterium of acute diarrhea. However, although there are patent applications for the detection kit and detection technology related to this bacterium (Application No.: 201210467910.1, title: "a double PCR detection primer set, kit and method for pathogenic snakehead-derived Aeromonas schubert ") disclosure, but the kit and detection technology of this application are only applicable ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/04C12Q1/686C12Q2531/113
Inventor 沈锦玉潘晓艺尹文林郝贵杰姚嘉赟徐洋蔺凌云袁雪梅
Owner ZHEJIANG INST OF FRESH WATER FISHERIES
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