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

Primer group and kit for synchronously detecting multiple tobacco viruses

A technology for simultaneous detection of tobacco virus, applied in the field of plant virology, can solve problems such as heavy workload and lack of test kits, and achieve the effect of reducing detection cost and improving detection efficiency

Inactive Publication Date: 2010-11-03
SHAANXI TOBACCO RES INST +1
View PDF1 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are only reports on the detection of TMV, CMV, etc. by single-plex PCR in China. However, in the field, it is often manifested as a compound infection of multiple diseases. The workload of using a single PCR to systematically detect multiple viruses is relatively large. The superiority in the detection of tobacco virus is more obvious, and there is currently a lack of kits that can simultaneously detect multiple tobacco viruses

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
  • Primer group and kit for synchronously detecting multiple tobacco viruses
  • Primer group and kit for synchronously detecting multiple tobacco viruses
  • Primer group and kit for synchronously detecting multiple tobacco viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1: Singleplex RT-PCR

[0058] Extraction of total viral RNA

[0059] Weigh 0.1 g each of tobacco diseased leaves (respectively marked as sample 2, sample 3, sample 4, sample 5, and sample 6) infected with five viruses of TMV, CMV, TEV, PVY, and TVBMV and freeze them at -80°C for grinding. In the bowl, add liquid nitrogen to fully grind, quickly transfer to an RNase-free sterile 1.5mL eppendof tube, add 1mL BIOzol Extraction Reagent to mix, and place for 15min. Add 0.2mL of chloroform and 0.3ml of phenol solution and invert up and down to mix well, and centrifuge at 12000×g for 15min at 4°C. Take the supernatant and transfer it to a new eppendof tube, add isopropanol equal to the volume of the supernatant, vortex and mix, and place at -20°C for 30 minutes. Centrifuge at 12000×g at 4°C for 15 minutes, and discard the supernatant. The precipitate was washed with 1 mL of 75% ethanol (prepared with DEPC water), centrifuged at 12000×g at 4°C for 5 min, the supernat...

Embodiment 2

[0066] Example 2: Multiplex RT-PCR

[0067] Extraction of total viral RNA

[0068] Take 6 samples respectively: healthy tobacco, 0.1 g each of TMV, TMV+CMV, TMV+CMV+TEV, TMV+CMV+TEV+PVY, TMV+CMV+TEV+PVY+TVBMV-infected tobacco leaves, 6 sample labels For sample A, sample B, sample C, sample D, sample E, sample F.

[0069] Place the samples in a deep-frozen mortar at -80°C, grind them thoroughly with liquid nitrogen, quickly transfer them to RNase-free sterile 1.5mL eppendof tubes, add 1mL BIOzol Extraction Reagent, mix well, and place for 15min. Add 0.2mL of chloroform and 0.3ml of phenol solution and invert up and down to mix well, and centrifuge at 12000×g for 15min at 4°C. Take the supernatant and transfer it to a new eppendof tube, add isopropanol equal to the volume of the supernatant, vortex and mix, and place at -20°C for 30 minutes. Centrifuge at 12000×g at 4°C for 15 minutes, and discard the supernatant. The precipitate was washed with 1 mL of 75% ethanol (prepared...

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 primer group and a kit for synchronously detecting multiple tobacco viruses. The kit consists of a component I and a component II, wherein the all the reagents in the component I and the component II are separately packed; the component I comprises reverse transcriptase, an RNA enzyme inhibitor, a dNTP mixture, a reverse primer sequence group, and reverse transcription reaction buffer; and the component II comprises PCR reaction buffer, a dNTP mixture, MgCl2, Taq DNA polymerase and a PCR primer group. The primer group is characterized in that: the reverse primer sequence group comprises SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8 and SEQ ID NO: 10; and the PCR primer group comprises SEQ ID NO: 1 to 10.

Description

technical field [0001] The invention belongs to the technical field of plant virology, in particular to a primer set and a kit for synchronous detection of multiple tobacco viruses. Background technique [0002] Tobacco is one of the most important economic crops in the world, and my country's planting area ranks first in the world. However, with the development of industry, the expansion of planting area and the adjustment of industrial structure, after the 1970s, the harm of tobacco virus disease has increased year by year. Due to the infection of the virus, the chlorophyll of tobacco is destroyed, which leads to the decline of photosynthesis, seriously affects the yield and quality of tobacco leaves, and causes great economic losses. [0003] The main diseases harmful to tobacco in my country are tobacco mosaic virus (Tobacco Mosaic Virus, TMV), cucumber mosaic virus (Cucumber Mosaic Virus, CMV), potato virus Y (Potato Virus Y, PVY), tobacco etch virus (Tobacco Etch Viru...

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): C12Q1/70C12Q1/68C12N15/11
Inventor 成巨龙吴云锋
Owner SHAANXI TOBACCO RES INST
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