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

Real-time fluorescent PCR detection method of L.maculans as well as primer and probe used for detection

A technology of real-time fluorescence and detection method, which is used in the detection of rape stem canker, the real-time fluorescence PCR detection of rape stem canker, the primers and probes for detecting rape stem canker

Inactive Publication Date: 2012-05-02
SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the real-time fluorescent PCR detection method using MGB probe to detect canker canker of rapeseed

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
  • Real-time fluorescent PCR detection method of L.maculans as well as primer and probe used for detection
  • Real-time fluorescent PCR detection method of L.maculans as well as primer and probe used for detection
  • Real-time fluorescent PCR detection method of L.maculans as well as primer and probe used for detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] In this embodiment, specific primers and Taqman-MGB probes are designed according to the specific site difference (G / A) of the rapeseed canker bacterium Leptosphaeria maculans and its close species ITS (internal transcribed spacer, the transcription spacer region of ribosomal gene) sequence The real-time fluorescent PCR technology was applied to the detection of L.maculans, the stem-base canker of rapeseed, and the experimental steps were as follows:

[0032] 1 Material method

[0033] 1.1 Test strains

[0034] Some strains of rapeseed canker L.maculans and rapeseed blackleg L.biglobosa used in the test were isolated and purified by our laboratory (Shanghai Entry-Exit Inspection and Quarantine Bureau) from imported rapeseed samples from Canada, Australia and Ukraine. Some strains were provided by Shenzhen, Jiangsu, and Ningbo Entry-Exit Inspection and Quarantine Bureau Technology Centers. The domestic rapeseed black shank L.biglobosa strain was provided by Mr. Li Qiang...

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

PropertyMeasurementUnit
pore sizeaaaaaaaaaa
Login to View More

Abstract

The invention discloses a real-time fluorescent PCR detection method of L.maculans as well as a primer and a probe used for detection. According to the difference of the sequences of the L.maculans and a similar species of ITS thereof, a primer Lmb3(5'-caccaattggatccccta-3') / R2(5'-ggcgcaaaa tgtgctgcgct-3') and a probe Probe-M(5'-FAM-cacttgggactcgccttg-MGB-3') which are used for detecting the L.maculans are designed, and the real-time fluorescent PCR detection method of the L.maculans is constructed. The sensitivity of the detection method reaches 4pg hypha DNA, the entire detection process is controlled within 4 hours, and the characteristics of high speed, specificity and sensitivity of the invention can meet the quick initial survey of an imported rapeseed sample and the quick identification of a pathogen isolate.

Description

technical field [0001] The invention relates to the technical fields of agriculture and plant quarantine, in particular to a method for detecting rapeseed canker, in particular to a real-time fluorescent PCR detection method for rapeseed canker. In addition, the invention also relates to primers and probes for detecting canker canker of rape. Background technique [0002] Rapeseed black shank (Phoma stem canker), also known as "rape black leg", is one of the most serious fungal diseases of rapeseed worldwide. The pathogen is Phoma stem canker, which mainly includes two species: Phoma stem canker Leptosphaeria biglobosa and Leptosphaeria maculans, two pathogenic bacteria, have similar morphological characteristics, but differ in their ability to cause stem infection and base ulcers in rapeseed. The former has weak pathogenicity and generally causes damage to the middle and upper parts of the stem, causing small losses. It is distributed in some rape-producing countries inclu...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11G01N21/64
Inventor 易建平尚琳琳周国梁
Owner SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
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