RPA (recombinase polymerase amplification) method for diagnosis and pathogen detection of blood flesh disease of watermelons

A pathogen and watermelon technology, which is applied in the RPA field of watermelon blood-flesh disease diagnosis and pathogen detection, to achieve the effects of shortening identification time, saving biochemical reagents, and reducing costs

Pending Publication Date: 2019-03-15
SHENYANG AGRI UNIV
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method basically meets the requirements for the detection of cucumber green

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
  • RPA (recombinase polymerase amplification) method for diagnosis and pathogen detection of blood flesh disease of watermelons
  • RPA (recombinase polymerase amplification) method for diagnosis and pathogen detection of blood flesh disease of watermelons
  • RPA (recombinase polymerase amplification) method for diagnosis and pathogen detection of blood flesh disease of watermelons

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0026] 1. CGMMV gene-specific primers

[0027] According to the genome sequence of CGMMV-lnxg isolate (KY040049.1), according to the design principles of RPA primers, the specific RPA primer pair CGMMV-F / CGMMV-R for detecting CGMMV was designed using software and manual correction. The primer sequences are shown in Table 1.

[0028] 2. Rapid extraction of total RNA from watermelon samples

[0029] Put 0.1 g of sample material into a cleaned and sterilized mortar and grind with liquid nitrogen, then add 1 mL of Trizol Reagent and mix well. Transfer the above homogenate sample to a 2ml RNase-free centrifuge tube, cap it, shake it vigorously and mix it evenly, and place it at room temperature for 5 minutes; add 0.2mL chloroform, mix it upside down for 15 seconds, and place it at room temperature for 15 minutes; 4°C, 12,000rpm, Centrifuge for 20min, transfer the upper colorless aqueous phase to a new 1.5ml RNase-free centrifuge tube; add an equal volume of isopropanol (about 400-...

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 RPA (recombinase polymerase amplification) method for diagnosis and pathogen detection of blood flesh disease of watermelons. An efficient and economic technical system for rapidly detecting a CGMMV (cucumber green mottle mosaic virus) is established with a technological means of modern molecular biology study, the RPA rapid detection method is provided, and the method comprises steps as follows: cDNA of a sample with the virus is taken as a template in a RPA reaction, a pair of detection primers are added, a reaction is performed for 20-30 min at constant temperaturebeing 37-40 DEG C in a water bath kettle, sensitivity reaches 10<-6> and is higher, the CGMMV is detected rapidly and accurately, meanwhile, identification time is shortened, biochemical reagents aresaved, cost is reduced, and theoretical basis and technical support are provided for monitoring, forecasting and control of the CGMMV as a quarantine disease.

Description

technical field [0001] The technical field of plant quarantine of the present invention relates in particular to an RPA method for the diagnosis of watermelon blood flesh disease and the detection of pathogens. Background technique [0002] Cucumber green mottle mosaic virus (CGMMV), a member of the Tobamovirus genus, is an important quarantine virus on cucurbit crops worldwide, seriously threatening watermelon, muskmelon, Production of Cucurbitaceae crops such as cucumbers. CGMMV can be transmitted by seeds, pollen, soil, water and insect vectors as well as mechanical contact. The leaves of CGMMV-infected plants will have obvious mosaic symptoms, growth retardation, and fruit deformity, which will eventually lead to a decrease in yield, resulting in huge economic losses. [0003] CGMMV has a variety of transmission methods, but the main ones are infected seeds and mechanical transmission. (1) Seed transmission: CGMMV is a typical seed-borne virus. The virus is mainly tra...

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): C12Q1/70C12Q1/6844
CPCC12Q1/6844C12Q1/701C12Q2521/507C12Q2522/101C12Q2563/107C12Q2531/119
Inventor 吴元华焦裕冰夏子豪蒋均匀
Owner SHENYANG AGRI UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap