Method for rapidly detecting pathogen of rice black-streaked dwarf

A technology for rice black-streaked dwarf and black-streaked dwarf disease, applied in biochemical equipment and methods, microbe measurement/inspection, etc., to achieve simple, fast, accurate and sensitive detection process

Inactive Publication Date: 2015-03-25
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This technology allows for quicker and more precise identification of different strains or types of bacteria by detecting specific DNA sequences called gene markers associated with them through amplified signals from two pairs of primer sets without requiring further analysis steps like enzymatic digestion. It can be used alone at home instead of relying on expensive equipment such as X ray fluorography (XRF).

Problems solved by technology

This patented problem addressed in this patents relates to improving the efficiency and accuracy of identifying various strains of harmful bacteria called RBS SDV during plant growth due to the development of efficient and reliable techniques for identification purposes. Current diagnostic tools like PCR cannot accurately differentiate between closely resided populations containing both RBS DWDSV and another susceptible variety.

Method used

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  • Method for rapidly detecting pathogen of rice black-streaked dwarf
  • Method for rapidly detecting pathogen of rice black-streaked dwarf
  • Method for rapidly detecting pathogen of rice black-streaked dwarf

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Experimental program
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Embodiment

[0020] Sample selection: The rice, corn and other samples of suspected diseased plants in the field were mainly collected from Zhejiang, Shandong, Hunan, Hainan and other provinces and cities in China between 2012 and 2014; the planthopper samples were mainly collected from various cities in Zhejiang Province.

[0021] Extraction of total RNA from samples: For plant samples such as rice, take an appropriate amount of leaves, add liquid nitrogen in a mortar, grind them into powder, transfer them to a 1.5mL centrifuge tube, and then add 1mL TRIzol reagent (Invitrogen); for planthopper samples, take a single Put the head parasites into a 1.5mL centrifuge tube, add 500 μL TRIzol reagent (Invitrogen), and then grind the parasites fully with a high-temperature-treated glass rod; extract total RNA according to the instructions of the TRIzol reagent for subsequent operations, and extract total RNA Store in -80°C refrigerator for later use.

[0022] Amplification reaction and melting curv

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Abstract

The invention relates to a method for rapidly detecting a pathogen of the rice black-streaked dwarf. The method comprises the following steps: performing RT-PCR amplification by using a pair of virus specific primers of a rice black-streaked dwarf virus and a southern rice black-streaked dwarf virus and by taking the total RNA of a sample to be detected as a template, and analyzing a high resolution solubility curve of an amplification product under the sealing condition, thereby judging the pathogen of the rice black-streaked dwarf. The method has the benefits that aiming at the production practice, a method for rapidly and synchronously detecting and distinguishing RBSDV from SRBSDV with high flux by using only one pair of the primers without any other probe or step of electrophoresis after amplification, EB dyeing or observation in ultraviolet lamps is developed and established, and technical support is provided for accurate diagnosis, prediction and forecasting and scientific prevention and control on the rice black-streaked dwarf.

Description

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Claims

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Application Information

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Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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