SYBR green I fluorogenic quantitative PCR (polymerase chain reaction) detecting method of pig proliferative enteropathy Lawsonia Intracellularis

A technology for Lawsonia intracellulare and porcine proliferative enteritis, applied in the field of bacterial detection, can solve the problems of reduced feed remuneration rate, complicated clinical symptoms, difficult to accurately judge, etc.

Inactive Publication Date: 2013-03-27
山东滨州沃华生物工程有限公司
View PDF1 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the disease has become a global epidemic. There are many recessively infected pigs, and the losses are easily overlooked. In recent years, China, Japan, South Korea and Taiwan in Asia have repeatedly occurred porcine proliferative enteritis. Sexual diarrhea, causing growth retardation of pigs, seriously reducing feed remuneration
In additi

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
  • SYBR green I fluorogenic quantitative PCR (polymerase chain reaction) detecting method of pig proliferative enteropathy Lawsonia Intracellularis
  • SYBR green I fluorogenic quantitative PCR (polymerase chain reaction) detecting method of pig proliferative enteropathy Lawsonia Intracellularis
  • SYBR green I fluorogenic quantitative PCR (polymerase chain reaction) detecting method of pig proliferative enteropathy Lawsonia Intracellularis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0033] (1) Primer design

[0034] According to the LI aspA gene (AY280626) registered in GenBank, a pair of primers were designed using Primer5.0 molecular biology software and sent to Shanghai Sangong Bioengineering Company for synthesis. The primer sequences are as follows:

[0035] PPE-1-F: 5--TAT, GGC, TGT, CAA, ACA, CTC, CG--3

[0036] PPE-1-R: 5--TGA, AGG, TAT, TGG, TAT, TCT, CC--3

[0037] The expected amplified fragment of PPE-1-F / R is 227bp.

[0038] (2) Extraction of DNA template

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 SYBR green I fluorogenic quantitative PCR (polymerase chain reaction) detecting method of pig proliferative enteropathy Lawsonia Intracellularis. The method provided by the invention comprises the following steps of: extracting pig Lawsonia Intracellularis DNA, designing primers, carrying out PCR amplification on aspa gene segments of PPE (polyphenylene oxide polyphenyl ether), cloning and identifying aspa genes of pig proliferative enteropathy, preparing a standard plasma template, amplifying fluorogenic quantitative PCR so as to obtain an amplification kinetic curve and a standard curve, testing the specificity, the sensitivity, the repeatability and stability, and detecting clinical disease material samples. The method provided by the invention has favorable specificity, sensitivity, repeatability and stability, and can identify and diagnose clinical suspected PPE infected or morbidity pigs simply, conveniently, rapidly, specifically, sensitively and accurately, and can detect and monitor the infection in the early stage.

Description

technical field [0001] The invention belongs to the technical field of bacterial detection, and in particular relates to a SYBR Green I fluorescent quantitative PCR detection method for Lawsonia intracellulare of porcine hyperplastic enteritidis. Background technique [0002] Porcine proliferative enteritis (PPE) is a contagious disease of pigs caused by Lawsonia intracellularis (LI). Lawsonia intracellulare is a curved obligate intracellular parasite, which mainly parasitizes the ileum, colon, and cecum of pigs and other animals, causing adenomatous hyperplasia of immature intestinal epithelial cells. At present, the disease has become a global epidemic. There are many recessively infected pigs, and the losses are easily overlooked. In recent years, China, Japan, South Korea and Taiwan in Asia have repeatedly occurred porcine proliferative enteritis. Sexual diarrhea, causing growth retardation of pigs, seriously reducing feed remuneration. In addition, the disease often h...

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): C12Q1/68C12Q1/04C12R1/01
Inventor 郭显坡陈申秒蔡培军
Owner 山东滨州沃华生物工程有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap