PCR detection method for detecting four kinds of poultry respiratory disease pathogens simultaneously

A detection method, respiratory technology, applied in the field of PCR detection of four kinds of poultry respiratory disease pathogens at the same time, can solve the problems of increasing experimental links and increasing experimental pollution, and achieve the effect of improving detection efficiency, simplifying operation steps, and improving accuracy

Active Publication Date: 2015-04-22
四川圣迪乐村生态食品股份有限公司 +1
View PDF4 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Due to the different reaction conditions, reverse transcription and PCR must be carried out in different reaction systems successively, increasing the number of experimental procedures and increasing the probability of experimental contamination
And because the optimal Tm val

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
  • PCR detection method for detecting four kinds of poultry respiratory disease pathogens simultaneously

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] (1) Extraction and purification of tissue total RNA: use the trizol method or a kit to extract the nucleic acid of the sample to be tested, and use it as the template used in the PCR reaction system;

[0042] (2) Design primers: According to the mRNA sequences in poultry tissues registered on Genbank, use Primer 5.0 to design primers for PCR detection of poultry gene expression, that is, design 4 pairs for the conserved regions of AIVH5, AIVH9, NDV and IBV genomes Primers:

[0043] AIVH5 upstream primer: 5ˊACAAAGCTCTCTATCAAAACCCAAC3ˊ

[0044] Downstream primer: 3ˊTACCATCTACCAACCATACCCAT5ˊ

[0045] AIVH9 upstream primer: 5ˊATCGGCTGTTAATGGAATGTGTT3ˊ

[0046] Downstream primer: 3ˊAATGGGATAAGTTCTGCGGGT5ˊ

[0047] NDV upstream primer: 5ˊGAGGTTACCTCYACYAAGCTRGAGA3ˊ

[0048] Downstream primer: 3ˊTCATTAACAAAYTGCTGCATCTTCCCWAC5ˊ

[0049] IBV upstream primer: 5ˊACTGAACAAAAGACAGACTTAGT3ˊ

[0050] Downstream primer: 3'CACTCTGAGCTGTTTGTGTTTG5'

[0051] On the prem...

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 belongs to the field of poultry disease detection and particularly relates to a one-step multiplex-PCR detection method for detecting four kinds of poultry respiratory disease pathogens simultaneously. The method is characterized by comprising the steps of providing a disease material possibly containing the four kinds of pathogens, extracting RNA, adding primers of the four kinds of pathogens to conduct PCR amplification and judging electrophoresis to obtain the result. The detection method is simple, convenient to operate and stable in accuracy and repeatability, whether the disease material contains the four kinds of pathogens can be confirmed in one experiment, 3/4 of detection cost is saved, detection time is shortened, and detection efficiency is improved.

Description

technical field [0001] The invention belongs to the field of poultry disease detection, in particular to a PCR detection method for simultaneously detecting four kinds of poultry respiratory disease pathogens. Background technique [0002] Avian influenza (Avian influenza, AI, including H5 subtype and H9 subtype), chicken Newcastle disease (Newcastle disease, ND), chicken infectious bronchitis (infectious bronchitis, IB) infection are four kinds that bring significant economic losses to the poultry industry. infectious disease. They have common respiratory symptoms clinically and can infect chickens, ducks, geese, pigeons and birds of different ages, causing high morbidity and mortality. Timely and accurate detection of these four pathogens is crucial for clinical disease prevention and control. [0003] Since the four pathogens are all RNA viruses, the traditional method requires a two-step reaction (reverse transcription and PCR) to obtain laboratory test results. The...

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): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/686C12Q2600/112C12Q2531/113C12Q2537/143
Inventor 邱荣斌赖守勋蒋清蓉王富妍代友洪刘旭
Owner 四川圣迪乐村生态食品股份有限公司
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