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

Application of riemerella anatipestifer Imp gene, PCR detection kit and method thereof

A Riemerella anatipestifer and detection kit technology, which is applied in the field of livestock and poultry detection, can solve the problems of unfavorable diagnosis of etiology, inspection of pathogenic conditions, confusion of Escherichia coli and Salmonella, and economic losses in the duck industry. The effect of simple result judgment and reduced testing cost

Active Publication Date: 2018-11-16
SICHUAN AGRI UNIV
View PDF4 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disease is prevalent in almost all duck raising areas in the world, causing huge economic losses to the duck industry
The traditional identification of RA usually detects phenotypic indicators such as its cultural characteristics, morphological staining, physiological and biochemical characteristics, and certain chemical compositions of the bacteria. to confuse
In addition, the traditional identification method is complicated to operate, time-consuming and laborious, which is not conducive to timely diagnosis of the cause, inspection of the pathogen and control of the spread of the disease
Aiming at these characteristics of Riemerella anatipestifer, many researchers have established some detection techniques, such as fluorescent antibody technique, ELLSA, immunohistochemistry, etc., which have been reported successively, but in practice, they all have different degrees of limitations, such as the need to target different serum type of antibody as a detection reagent

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
  • Application of riemerella anatipestifer Imp gene, PCR detection kit and method thereof
  • Application of riemerella anatipestifer Imp gene, PCR detection kit and method thereof
  • Application of riemerella anatipestifer Imp gene, PCR detection kit and method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Embodiment 1, the establishment of Riemeria anatipestifer PCR method

[0019] Design of primer pairs for specific amplification of R. anatipestifer: The Imp gene was found from the genomic DNA sequence of R. anatipestifer through bioinformatics analysis. Imp is highly conserved in other Gram-negative bacteria and has multiple Important biological functions, in addition to participating in the process of lipopolysaccharide export and assembly to the outer membrane, are also related to the formation of bacterial envelope and the tolerance of bacterial organic solvents. It is used as the detection target gene of Riemerella anatipestifer, and the gene sequence is shown in SEQ ID NO.1.

[0020] (1) Primer design

[0021] Input the DNA sequence of this gene into the primer design software Primer Premier 5.0 to design primers, set the product size range to 100-500bp, select primers from the alternative primer pair, and the primer sequences are as follows (primers were provide...

Embodiment 2

[0031] Embodiment 2, PCR detection method specificity evaluation of Riemeria anatipestifer

[0032] According to the DNA template extraction method and PCR detection method in Implementation 1, Escherichia coli, Pasteurella, Salmonella, and various serotypes of Riemeria anatipestifer strains (reserved strains or serological tests identified as Riemerella anatipestifer The clinical isolates, as shown in Table 1) were tested.

[0033] Table 1. The strains used for specificity evaluation and the detection results of PCR test

[0034] Strain number

Deposit number or clinically isolated and identified strains

Strain name and serotype

PCR result

RA CH-1 (RCH)

CCTCC NO.M2017702

Riemerella anatipestifer serotype 1

+

RA CH-2

Identification of serotyped clinical isolates

Riemerella anatipestifer serotype 2

+

RCAD0166

Identification of serotyped clinical isolates

Riemerella anatipestifer serotype 2

+

RA ATC...

Embodiment 3

[0037] Example 3. Sensitivity evaluation experiment of Riemeria anatipestifer PCR detection method

[0038]The Riemeria anatipestifer template DNA obtained by extracting in Example 1 was measured by OD260 / 280, and the concentration of the Riemeria anatipestifer total DNA solution was 150 ng / μl, and was diluted 10 times with sterile water, and co-diluted 8 gradients, 2 μL of each gradient was added to the PCR reaction system, and the DNA template was subjected to PCR amplification detection according to the method in step (3) of Example 1. Take 5 μL of the PCR amplification product, and perform electrophoresis in 1% agarose gel, and observe the gel electrophoresis result in a gel imager as follows: figure 2 shown. Depend on figure 2 It can be seen that a clear band can be seen in the fifth lane, corresponding to a DNA concentration of 15 pg, which has extremely high sensitivity.

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 relates to a PCR detection kit based on riemerella anatipestifer Imp gene and a method thereof. The kit contains primer pairs shown as SEQ ID NO.2 and SEQ ID NO.3, wherein the nucleotidesequence of the riemerella anatipestifer Imp gene is shown as SEQ ID NO.1. The detection kit and method provided by the invention adopt Imp gene as the target gene to detect riemerella anatipestifer,have significant specificity and high sensitivity, and are of important significance for early detection of riemerella anatipestifer.

Description

technical field [0001] The invention belongs to the field of livestock and poultry detection, relates to the application of the Riemerella anatipestifer Imp gene, and also relates to a PCR detection kit based on the Riemerella anatipestifer Imp gene and a corresponding detection method. Background technique [0002] Riemerella anatipestifer (RA) is the pathogenic bacterium of duck infectious serositis, which is one of the important diseases that endanger the duck industry. The disease is an acute or chronic sepsis process, characterized by fibrinous pericarditis, perihepatitis, air sac inflammation, meningitis, and some caseous salpingitis and arthritis. The morbidity rate is 10%-90%, and the case fatality rate is as high as 80%, which can occur throughout the year. The disease is prevalent in almost all duck raising areas in the world, causing huge economic losses to the duck industry. The traditional identification of RA usually detects phenotypic indicators such as its ...

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): C12Q1/689C12Q1/686C12Q1/04C12N15/11
CPCC12Q1/686C12Q1/689C12Q2565/125
Inventor 汪铭书邱浩程安春
Owner SICHUAN AGRI UNIV
Features
  • Generate Ideas
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com