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

Two step method multiple RT-PCR solidified detection reagent kit for potato virus disease and its detecting method

A potato virus and RT-PCR technology, applied in the field of agricultural biology, can solve the problems of difficulty in ensuring the quality of qualified seedlings, lack of positive controls, and restrictions on the potential of potato virus-free seedlings to increase production.

Inactive Publication Date: 2006-11-08
CHONGQING UNIV +2
View PDF0 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Aiming at the current potato virus molecular diagnostic kits that are not produced in China, and a small number of imported serological kits have poor specificity, lack of positive controls, and inability to determine the type of virus, etc., it is difficult to guarantee the quality detection technology of qualified seedlings, which seriously restricts the potato virus. Application of Poisonous Seed Potato and Its Yield-increasing Potential

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
  • Two step method multiple RT-PCR solidified detection reagent kit for potato virus disease and its detecting method
  • Two step method multiple RT-PCR solidified detection reagent kit for potato virus disease and its detecting method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0134] Example 1. Two-step duplex RT-PCR simultaneous detection of potato virus Y and potato leafroll virus solid-phase detection kit and detection method.

[0135] 1. Components of the solid-phase detection kit:

[0136] Sample virus RNA extraction reagent, PVY, PLRV viral RNA reverse transcription solid-phase reagent, cDNA amplification solid-phase reagent, harmless PVY, PLRV positive control, healthy plant negative control.

[0137] 2. Preparation of solid-phase kit:

[0138] 1. The detection method includes the following steps:

[0139] 1) Extracting viral RNA: preparing potato virus RNA special extract solution (0.5% Triton X-100 and containing final concentration 0.4% NaSO 3 Mixed solution with an appropriate amount of purified water without RAN enzyme at room temperature); take out the potato sample to be tested, and add 10 mg of leaves, 30 mg of leaf stems, 40 mg of stems or 60 mg of tubers of the sample potato plants into 100 μL of potato virus RNA extract Extract ...

Embodiment 2

[0147] Example 2. Two-step triple RT-PCR simultaneous detection of Potato virus X, Potato virus A and Potato S virus solid-phase detection kit and detection method.

[0148] 1. Components of the solid-phase detection kit:

[0149] Sample viral RNA extraction reagents, PVX, PVA, PVS viral RNA reverse transcription solid-phase reagents, cDNA amplification solid-phase reagents, harmless PVX, PVA, PVS positive controls, healthy plant negative controls.

[0150] 2. Preparation of solid-phase kit:

[0151] 1. The detection method includes the following steps:

[0152] 1) extract viral RNA: prepare potato virus RNA special extraction liquid (being 0.5% Triton X-100 and containing final concentration 0.4% NaSO 3 Mixed solution with an appropriate amount of purified water without RAN enzyme at room temperature); take out the potato sample to be tested, and add 5 mg of leaves, 20 mg of leaf stems, 30 mg of stems or 50 mg of tubers of the sample potato plants into 100 μL of potato viru...

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 present invention relates to two-step synchronous double RT-PCR process of detecting Y virus and leaf roll virus of potato, synchronous triple RT-PCR process of detecting X virus, A virus and S virus of potato, and the detecting kit. The present invention can detect five main potato viruses within 4 hr in high sensitivity, high specificity, high stability and high reliability. The present invention is suitable for the early diagnosis on potato seedling and potato seeds, and is significant on the quality monitoring of potato seed and virus disease prevention.

Description

technical field [0001] The invention relates to an agricultural biotechnology, in particular to a simple and rapid solid-phase nucleic acid detection kit for synchronously amplifying multiple potato viruses, which is suitable for use in departments such as plant protection and agricultural product quality technical supervision. Background technique [0002] Potato is a high-yield crop worldwide, and my country's planting area and output rank first in the world. Potato virus disease is an important restrictive factor in potato production and the main cause of potato variety degeneration, seriously affecting the yield and quality of potato. In my country, several main viruses and viroids that harm potatoes are Potato Virus X (PVX for short), Potato Virus Y (PVY for short), Potato Leafroll Virus (PLRV for short), Potato Virus A (PVA for short), Potato Virus S (PVS for short), wherein the most serious virus is PLRV, PVY. In potato production, seve...

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 王中康殷幼平袁青夏玉先彭国雄曾德玉曹月青刘红光
Owner CHONGQING 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