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

Kit and method for detecting virus infecting situation of Chinese giant salamander colony iridovirus by use of ecdysis

A technology of iridescent virus and kit, applied in the field of kit and method for detecting iridescent virus in giant salamander group by using molting

Inactive Publication Date: 2011-04-27
NORTHWEST A & F UNIV +1
View PDF3 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] By randomly obtaining fresh molting from the giant salamander population without harming any individual in the giant salamander population, it overcomes the problem of obtaining samples from the visceral tissue of giant salamanders

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
  • Kit and method for detecting virus infecting situation of Chinese giant salamander colony iridovirus by use of ecdysis
  • Kit and method for detecting virus infecting situation of Chinese giant salamander colony iridovirus by use of ecdysis
  • Kit and method for detecting virus infecting situation of Chinese giant salamander colony iridovirus by use of ecdysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Trial production of a giant salamander iridescent virus detection kit

[0045] (Specification: test 10 samples according to one kit)

[0046] Solution A, molting sample lysate, 1 tube of 12ml, mainly containing PBS; 1% triton-100

[0047] Solution B, 1 tube 400ul, 200ug / ml proteinase K;

[0048] Solution C, 1 tube, template extract, 1 tube 12ml, containing phenol / chloroform / isoamyl alcohol, the ratio is 25:24:1;

[0049] Solution D, 1 tube 2ml, 3mol / l sodium acetate;

[0050] Solution E, 1 tube of 20ml, containing absolute ethanol;

[0051] Solution F, 1 tube 40ml, containing 75% ethanol;

[0052] Tube G, 1ml per tube, containing sterilized ultrapure water;

[0053] Solution H, S1 tube 300μl, containing PCR amplification reaction solution (20μl system), including deionized water, dNTP substrate, containing Mg 2+ 1× buffer; Taq enzyme; primer S1 (5’-CCCCTCCCATTCTTTCTTCTCC-3’ and

[0054] 5'-GGCGTTGGTCAGTCTACCGTAAT-3').

[0055]S2 tube 300μl, containing PCR amplif...

Embodiment 2

[0062] Detect 10 giant salamander group iridescent virus carrying situations with above-mentioned kit of the present invention:

[0063] 1. According to the requirements of the present invention, obtain 10 different groups of giant salamander moults, each of which has 5 slices, respectively cut 10-20 times with sampling scissors in a 2 μl centrifuge tube, and mix well.

[0064] 2. Put 0.1g of molting sample into a new 1.5ml centrifuge tube numbered (No. 3-12), add 500μl of solution A, freeze and thaw 3 times repeatedly, add 20μl of solution B, bathe in 55℃ water for 1.0h, and centrifuge at 4000rpm for 10min , to molt debris.

[0065] 3. Take 500 μl of the supernatant into a new 1.5ml centrifuge tube (corresponding to the number in the previous step), add an equal volume (500 μl) of C solution for extraction, mix well, place at room temperature for 5 minutes, and centrifuge at 12,000 rpm for 5 minutes;

[0066] 4. Take the supernatant into a new 1.5ml centrifuge tube (correspo...

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 discloses a kit and for detecting virus infecting situation of Chinese giant salamander colony iridovirus by use of ecdysis. The kit comprises 1 tube of solution A, namely ecdysis sample lysis solution containing PBS (phosphate buffer saline) and 1% triton-100, 1 tube of solution B containing 200mug / ml of protease K, 1 tube of solution C, namely template extraction solution containing phenol, chloroform and isoamyl alcohol based on the proportion of 25:24:1, 1 tube of solution D containing 3mol / l sodium acetate and the like. In the invention, the fresh Chinese giant salamander ecdysis is taken as a pathological material detection sample, thus solving the defect that the detection sample is obtained by killing Chinese giant salamander individuals; according to the characteristics of the Chinese giant salamander ecdysis, the method for extracting virus genome DNA from the ecdysis is optimized, and the kit for simply and effectively extracting an iridovirus gene from the Chinese giant salamander ecdysis sample as well as 2 pairs of PCR (polymerase chain reaction) primers for 2 conserved genes of the iridovirus and PCR conditions are designed; and positive and negative reference substances are established. By utilizing the kit for detection in standard steps, the virus infecting situation of the Chinese giant salamander colony iridovirus can be more reliably mastered, thus being favorable for application to actual production.

Description

technical field [0001] The invention relates to a kit and a method for detecting the poisonous condition of giant salamander group iridescent virus by using molting. Background technique [0002] The distribution of giant salamanders in the world is mainly in Japan, North America and my country. In terms of resource protection and industrial development of giant salamanders, although Japanese giant salamanders and American cryptobranch salamanders have paid attention to resource protection to varying degrees, my country has made great efforts in research on giant salamander domestication and breeding techniques and Its development and utilization are already in the leading position in the world. The artificial breeding of giant salamanders has not only become a means of ecological environment protection, but also has become a characteristic breeding industry in some areas. Artificial breeding of giant salamander has great ecological and economic value. [0003] Chinese giant...

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 董武子杨长明张晓明邢福珊覃金洲胡沈荣宋峰峰安俊辉张驰
Owner NORTHWEST A & F 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