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

A segment of sequence and a restriction enzyme site for differentiating canine distemper viral vaccine strain and wild strain

A technology of canine distemper virus and enzyme cutting site, applied in the direction of microbial measurement/testing, biochemical equipment and methods, etc., can solve the problems of no treatment and comprehensive prevention and control, difficult to determine, etc., achieve high accuracy and efficiency, The effect of the simple identification method

Inactive Publication Date: 2009-03-04
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
View PDF0 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the vaccine has a certain half-life in the animal body, it is difficult to determine whether the detected canine distemper virus is the residual of the vaccine or infected with wild virus when the vaccine is immunized in the early stage of the disease, so there is no better treatment and comprehensive prevention and control measures.

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
  • A segment of sequence and a restriction enzyme site for differentiating canine distemper viral vaccine strain and wild strain
  • A segment of sequence and a restriction enzyme site for differentiating canine distemper viral vaccine strain and wild strain

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] 1. Design and synthesis of primers:

[0011] A pair of primers P1: 5'TTCTG AGGCA GATGA GTTCT TC3', P2: 5'CTTGGATGCT ATTTC TGACA CT 3' were designed and synthesized, and the target fragment size to be amplified is expected to be 829bp. The fragment of CDV NP gene 829 in the vaccine strain CDV3 strain of the Specialty Research Institute was amplified.

[0012] 2. Viral RNA preparation RNA was extracted using Trizol reagent from Invitrogen Company. The main steps are: (1) aseptically take the viscera or blood of the diseased animal, and add 1:1 PBS to the viscera for grinding; (2) take 250 μl of the grinding suspension, add 750 μl Trizol and 200 μl chloroform successively, shake vigorously for 15 seconds, Centrifuge at 14000rpm at 4°C for 15min. (3) Take out the supernatant and transfer it to a new centrifuge tube, add an equal volume of isopropanol, mix evenly by inversion, settle at room temperature for 10 minutes, and centrifuge at 14000 rpm at 4°C for 10 minutes. (4...

Embodiment 2

[0021] 1. Design and synthesis of primers:

[0022] A pair of primers P1: 5'TTCTG AGGCA GATGA GTTCT TC 3', P2: 5'CTTGGATGCT ATTTC TGACA CT 3' were designed and synthesized, and the expected amplified target fragment size is 829bp. The 829 fragment of CDV NP gene in the Ondersterpoort strain from the joint seedlings of the French Vic company was amplified.

[0023]2. Viral RNA preparation RNA was extracted using Trizol reagent from Invitrogen Company. The main steps are: (1) Aseptically take the viscera or blood of the diseased animal, and add 1:1 PBS to the viscera for grinding; (2) Take 250 μl of grinding suspension and add 750 μl Trizol and 200 μl chloroform successively, shake vigorously for 15 seconds, Centrifuge at 14000rpm at 4°C for 15min. (3) Take out the supernatant and transfer it to a new centrifuge tube, add an equal volume of isopropanol, mix evenly by inversion, settle at room temperature for 10 minutes, and centrifuge at 14000 rpm at 4°C for 10 minutes. (4) P...

Embodiment 3

[0033] 1. Design and synthesis of primers:

[0034] A pair of primers P1: 5'TTCTG AGGCA GATGA GTTCT TC 3', P2: 5'CTTGG ATGCTATTTC TGACA CT3' were designed and synthesized, and the expected amplified target fragment size is 829bp. The fragment of CDVNP gene 829 from the Hebei Changli raccoon dog HCL07 strain was amplified.

[0035] 2. Viral RNA preparation RNA was extracted using Trizol reagent from Invitrogen Company. The main steps are: (1) Aseptically take the viscera or blood of the diseased animal, and add 1:1 PBS to the viscera for grinding; (2) Take 250 μl of grinding suspension and add 750 μl Trizol and 200 μl chloroform successively, shake vigorously for 15 seconds, Centrifuge at 14000rpm at 4°C for 15min. (3) Take out the supernatant and transfer it to a new centrifuge tube, add an equal volume of isopropanol, mix evenly by inversion, settle at room temperature for 10 minutes, and centrifuge at 14000 rpm at 4°C for 10 minutes. (4) Pour off the liquid, add 1ml of 75...

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 method used to distinguish a sequence and a restriction enzyme cutting site of a canine distemper virus vaccine strain and a field strain and integrate two molecular biology test methods, namely RT-PCR and RFLP; a CDV NP gene is selected; a special primer is designed to amplify 829bp, and the segment is treated by BamHI RFLP; through agarose gel electrophoresis judgment, the canine distemper virus tested in the animal body is identified to belong to an inoculated vaccine strain or be injected by field virus from the number of segment; two segments 675bp and 153bp obtained from the electrophoresis are vaccine strains, and three segments 455bp, 220bp and 153bp are field strains. The method effectively distinguishes the canine distemper virus vaccine strain and the field strain, and provides veterinary technical guarantee for the cultivation of fur animals.

Description

Technical field: [0001] The invention relates to a sequence and an enzyme cutting site for distinguishing canine distemper virus vaccine strains from wild strains, which are fusion reverse transcription-polymerase chain reaction (RT-PCR) and restriction endonuclease analysis ( RFLP) the invention of two conventional molecular biology identification detection methods, select specific nucleic acid fragments and a restriction endonuclease, effectively distinguish the vaccinated strain and the infected wild strain of canine distemper virus. Background technique: [0002] Neutralization test, immunofluorescence antibody technique, enzyme-linked immunosorbent assay and other methods have played a role in the diagnosis of canine distemper. In addition, compared with the above methods, the detection rate of the electron microscope diagnosis method has a significant consistency. Although the above methods have achieved certain effects, some of them take a long time, and some have po...

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): C12Q1/68
Inventor 王凤雪闫喜军吴威柴秀丽张海玲罗国良邵西群
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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