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

Applications of echinococcus granulosus mitochondrion gene ND6

A technology of Echinococcus granulosus and Echinococcus granulosus, applied in the application field of Echinococcus granulosus mitochondrial ND6 gene, achieving high reliability and improving detection efficiency

Active Publication Date: 2018-12-28
SICHUAN AGRI UNIV
View PDF5 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, slaughterhouses usually only rely on visual observation to make judgments without histological identification, and the judgment error rate can be as high as 15.4%.

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
  • Applications of echinococcus granulosus mitochondrion gene ND6
  • Applications of echinococcus granulosus mitochondrion gene ND6
  • Applications of echinococcus granulosus mitochondrion gene ND6

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Primer Design

[0027] According to the ND6 gene sequence of the complete mitochondrial genome of Echinococcus granulosus (sequence number: KJ559023.1) in GenBank, a pair of primers was designed using Primer Premier 5.0 software. upstream primer A 1 : 5'-TTTCGTGCTGTAGATGGT-3' (shown in SEQ ID NO: 1), downstream primer A 2 : 5'-CACAGATTTCAAAGGGTT-3' (shown in SEQ ID NO: 2), the amplified fragment is 558bp (complete ND6 wild-type gene 456bp and its upstream partial tRNA-GLU sequence 38bp, downstream partial tRNA-TYR sequence 64bp Composition, the sequence is shown in SEQ ID NO:5). The primer sequences were synthesized by Biotechnology (Shanghai) Co., Ltd.

Embodiment 2

[0028] Embodiment 2: PCR amplification

[0029] The amplification system is 20 μL: 2×Taq PCR MasterMix 10 μL, Primer A 1 、A 2 0.5 μL each, 4.0 μL template DNA, ddH 2 O 5 μL, mix gently and centrifuge briefly for 15s, at the same time, add ddH 2 O was used instead of template DNA as a blank control. Amplification conditions: pre-denaturation at 94°C for 3min; denaturation at 94°C for 10s, annealing at 58°C for 20s, extension at 72°C for 30s, a total of 30 cycles; final extension at 72°C for 5min. After the reaction, 10 μL of the amplified product was identified by 1% agarose gel electrophoresis. Parallel experiments were repeated 3 times.

Embodiment 3

[0030] Example 3: PCR amplification of Echinococcus granulosus cyst DNA

[0031]Echinococcus granulosus cysts were enzymatically hydrolyzed using Animal Tissue Direct PCR Kit (Foregene, Chengdu, China). Add 100μl Buffer AL and 4μl Foregene Protease to a 2ml centrifuge tube, and vortex slightly to mix. Put 5-10 mg of the treated cyst into the centrifuge tube, vortex slightly to mix, incubate at 65°C for 20 minutes, treat at 95°C for 5 minutes, and centrifuge at 12,000 rpm for 5 minutes. Then transfer the supernatant to a new centrifuge tube and store it at -20°C for later use or use it directly.

[0032] with synthetic primer A 1 、A 2 , using the above sample as a template, amplified according to the method of Example 2, the results are shown in figure 1 , the size of the band is in line with the expected result, and the band is clear, and there is no band in the negative control; in addition, after gene sequencing by the reagent company, the result is in line with expectat...

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 the technical field of biology and discloses a series of new applications of an echinococcus granulosus mitochondrion gene ND6. The echinococcus granulosus mitochondrion geneND6 is taken as molecular diagnosis marker, whether a slaughtered animal is infected with echinococcus granulosus or not is detected by PCR, high reliability, specificity and sensitivity are achieved,meanwhile, by means of direct PCR, detection efficiency can be improved, and the invention is particularly suitable for large-scale gene detection and high-throughput analysis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of the Echinococcus granulosus mitochondrial ND6 gene. Background technique [0002] The middle-stage larvae (Echinococcus granulosus) of Echinococcus granulosus mainly parasitize the liver and lungs of humans and animals (cattle, sheep, pigs, etc.), causing echinococcosis (also known as echinococcosis) in humans and animals. echinococcosis). Echinococcus granulosus is a zoonotic parasitic disease with worldwide distribution, and China is one of the areas with high incidence of echinococcosis. The disease has caused serious global economic and public health problems. It is estimated that global human cystic echinococcosis loses at least 1-3.6 million disability-adjusted life years (DALYs) per year, while the annual economic loss of animal cystic echinococcosis is at least 2 billion US dollars, of which China The loss of animal husbandry even reached 1.6 billion yua...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/158
Inventor 杨光友詹佳飞
Owner SICHUAN AGRI UNIV
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