RPA (recombinase polymerase amplification) primer and kit for quickly detecting citrus huanglongbing Asian species as well as detection method and application of RPA primer and kit

A citrus huanglongbing and detection kit technology is applied in the field of RPA primers for rapid detection of citrus huanglongbing in Asian species, which can solve the problems of citrus huanglongbing pathogen detection reports and no RPA technology, and achieve sustainable development and reduce false positives. The probability of positive, the effect of eliminating the interference of false positives

Pending Publication Date: 2021-01-29
PLANT PROTECTION RES INST OF GUANGDONG ACADEMY OF AGRI SCI
View PDF5 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, RPA technology has been used in the rapid diagnosis of medical pathogens, and there are some reports on the d

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) primer and kit for quickly detecting citrus huanglongbing Asian species as well as detection method and application of RPA primer and kit
  • RPA (recombinase polymerase amplification) primer and kit for quickly detecting citrus huanglongbing Asian species as well as detection method and application of RPA primer and kit
  • RPA (recombinase polymerase amplification) primer and kit for quickly detecting citrus huanglongbing Asian species as well as detection method and application of RPA primer and kit

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0042]Example 1

[0043](1) Experimental process

[0044](1) Preparation of sample template: Take about 0.2g of fresh leaves of the test plant, and extract the total plant DNA according to the operation steps of the Plant Genomic DNA Extraction Kit (EasyPure Plant Genomic DNA Kit, Beijing Quanshijin Biotechnology Co., Ltd.). Store in the refrigerator at -20℃ for later use.

[0045](2) RPA primer design and screening: Based on the 16S rDNA conservative sequence of Candidatus Liberibacterasiaticus (GenBank sequence accession number: AY192576.1), the primer HBas-F / HLBas-R was designed according to the primer requirements of RPA; The length is 397bp.

[0046]HLBas-F: 5'-CTTTAATACTGGTTGTCTAGAGTTTAGGAGAG-3',

[0047]HLBas-R: 5'-GAAAAGAAAATACCATCTCTGATATCGTCCTATA-3'.

[0048]Based on the OMP conservative sequence of the Asian species of Citrus Huanglongbing (GenBank sequence accession number: AY842429.1), two pairs of primers were designed according to the primer requirements of RPA: OMP-F1 / R1, OMP-F2 / R2; t...

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 an RPA (recombinase polymerase amplification) primer and a kit for rapidly detecting citrus huanglongbing Asian species as well as a detection method and application of the RPAprimer. The RPA amplification primer HLBa-F/HLBa-R developed on the basis of the conserved sequence 16S rDNA of the citrus huanglongbing Asian species establishes an RPA detection method for detecting the citrus huanglongbing Asian species for the first time, effectively eliminates false positive interference, is short in detection time consumption, economical and applicable, does not need expensive instruments and equipment such as a PCR instrument and a fluorescent quantitative PCR instrument, and is simple and easy to learn; powerful technical support can be provided for early detection ofcitrus yellow shoot of disease-free citrus seedlings in China, detection and confirmation of suspected citrus yellow shoot trees in fields and popularization of the detection capability of grass-roots scientific research units, so healthy development of disease-free citrus seedling breeding in China is promoted, spreading and diffusion of citrus yellow shoot in wide citrus production areas are early warned in advance, and the early warning effect is achieved; sustainable development of the citrus industry in China is further promoted.

Description

technical field [0001] The invention relates to a detection technology of plant quarantine diseases in my country, in particular to an RPA primer, a kit, a detection method and application thereof for rapidly detecting citrus huanglongbing Asian species. Background technique [0002] Citrus Huanglongbing (Huanglongbing) is a worldwide destructive disease, which has the characteristics of strong outbreak, rapid development, and great harm. It is listed as a quarantine disease in my country. Citrus huanglongbing is spreading in citrus growing areas all over the world, such as Asia, Africa, America and other regions. Citrus greening disease has a long duration, wide range and high incidence rate, resulting in short lifespan, low yield, high production cost and heavy economic losses of citrus in some areas of my country, which seriously restricts the healthy development of my country's citrus industry. In 2017, the area of ​​citrus Huanglongbing disease in the 10 provinces (dis...

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): C12Q1/689C12Q1/6844C12Q1/04C12N15/11C12R1/01
CPCC12Q1/689C12Q1/6844C12Q2521/507C12Q2565/125C12Q2521/101C12Q2522/101
Inventor 宋晓兵彭埃天黄峰凌金锋崔一平陈霞
Owner PLANT PROTECTION RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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