Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Primers for quickly detecting Edwardsiella tarda by using gene amplification method

A technology for Edwardsiella tarda and gene amplification, which is applied in the field of primers for rapid detection of Edwardsiella tarda by gene amplification method, and can solve the problems that cannot be popularized and applied

Active Publication Date: 2012-07-11
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
View PDF0 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above methods have the advantages of rapidity and specificity compared with the traditional routine identification of bacteria. However, the above methods are restricted by the preparation of antigens and antibodies, and the need for special equipment such as PCR instruments and gel electrophoresis. They cannot be widely used in production.
However, there is no relevant research report on the detection of E.tarda by using 6 primers to detect E.tarda by gene isothermal amplification technology, and there is no report on the detection of E.tarda by this technology in China.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Primers for quickly detecting Edwardsiella tarda by using gene amplification method
  • Primers for quickly detecting Edwardsiella tarda by using gene amplification method
  • Primers for quickly detecting Edwardsiella tarda by using gene amplification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0156] Embodiment 1: Carry out bacterial specific detection with the gene amplification primer of E.tarda

[0157] Firstly, the primers provided by the present invention are synthesized, and then the specific detection of E.tarda is carried out according to the following steps.

[0158] (1) Preparation of bacterial DNA: The DNA of bacterial samples was extracted using TIANamp Bacteria DNA Kit from Tiangen Biochemical Technology (Beijing) Co., Ltd., and the extracted DNA was used on DNA agarose gel Recovery Kit (Zymo, USA) was used for purification. After the DNA sample was prepared, it was placed at 95°C for 5 minutes, and then denatured in an ice bath for 5 minutes before being used as a template.

[0159] Prepare the reaction system: 0.2 μM each of primer 1 and primer 2 of the first set of primers, 1.6 μM each of primer 3 and primer 4, 0.8 μM each of primer 5 and primer 6, 1.4 mM each of dNTP, MgCl 2 6mM, Betaine 1.2M, Tris-HCl 20mM, KCl 10mM, MgSO 4 2mM, (NH 4 ) 2 SO...

Embodiment 2

[0164] Embodiment 2: Carry out bacterial specific detection with the gene amplification primer of E.tarda

[0165] First, the primers provided by the present invention are synthesized, and then the bacteria-specific detection of E. tarda is carried out according to the following steps.

[0166] (1) Preparation of bacterial DNA: The DNA of bacterial samples was extracted using TIANamp Bacteria DNA Kit from Tiangen Biochemical Technology (Beijing) Co., Ltd., and the extracted DNA was used on DNA agarose gel Recovery Kit (Zymo, USA) was used for purification. After the DNA sample was prepared, it was placed at 95°C for 5 minutes, and then denatured in an ice bath for 5 minutes before being used as a template.

[0167] Prepare the reaction system: 0.2 μM each of primer 1 and primer 2 of the fourth set of primers, 1.6 μM each of primer 3 and primer 4, 1.4 mM each of dNTP, MgCl 2 6mM, Betaine 1.2M, Tris-HCl 20mM, KCl 10mM, MgSO 4 2mM, (NH 4 ) 2 SO 4 10 mM, Triton X-100 0.1%...

Embodiment 3

[0172] Embodiment 3: Carry out bacterial specific detection with the gene amplification primer of E.tarda

[0173] First, the primers provided by the present invention are synthesized, and then the bacteria-specific detection of E. tarda is carried out according to the following steps.

[0174] (1) Preparation of bacterial DNA: DNA of bacterial samples was extracted using TIANamp Bacteria DNA Kit from Tiangen Biochemical Technology (Beijing) Co., Ltd. Recovery Kit (Zymo, USA) was used for purification. After the DNA sample was prepared, it was placed at 95°C for 5 minutes, and then denatured in an ice bath for 5 minutes before being used as a template.

[0175] Prepare the reaction system: 0.2 μM each of primer 1 and primer 2 of the fifth set of primers, 1.6 μM each of primer 3 and primer 4, 0.8 μM each of primer 5 and primer 6, 1.4 mM each of dNTP, MgCl 2 8mM, Betaine 1.2M, Tris-HCl 20mM, KCl 10mM, MgSO 4 2mM, (NH 4 ) 2 SO 410 mM, Triton X-100 0.1%, Bst DNA polymerase...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses gene amplification primers for quickly detecting Edwardsiella trada. Specific primers are designed by taking esrB of E. tarda or a hlyB upstream sequence of the Edwardsiella trada as a target. A gene amplification detection method by the primers has the characteristics of simplicity, convenience, quickness, specificity and sensitivity; genes with 10 copy number can be detected in 40 minutes only through a water bath or a metal bath; and the method is suitable for quickly detecting the E. tarda in field and has good application prospect.

Description

technical field [0001] The invention belongs to the technical field of microorganism detection, in particular to a primer for rapid detection of Edwardsiella tarda by a gene amplification method and an application thereof. Background technique [0002] E. tarda is a Gram-negative short bacillus, Edwardsiella genus, first reported by Hoshina in 1962, and it is one of the pathogenic bacteria with great harm in the current aquaculture industry. The host range of the bacterium is very wide. In addition to infecting a variety of fish, it has also been reported to infect amphibians, reptiles and mammals (including humans). Therefore, in order to diagnose, monitor and prevent E.tarda in time, it is of great significance to establish a fast, accurate and simple E.tarda detection technology. [0003] Conventional bacterial identification methods include the identification of pathogenic morphology, physiological and biochemical levels, and protein levels. However, due to the shortcom...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/68C12N15/11C12R1/01
Inventor 黄倢谢国驷张庆利史成银王秀华杨冰刘莉
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com