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

Primer probe composition for detecting pythium oligandrum, kit, application and detection method

A primer probe, Pythium technology, applied in the field of genetic engineering, can solve the problems of long cycle, poor specificity and low sensitivity of detection methods, and achieve the effects of increasing application value, good practicability and cumbersome operation.

Active Publication Date: 2022-03-11
NANYANG NORMAL UNIV
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the problems of long period, poor specificity and low sensitivity of the Pythium oligandrum biological detection method in the prior art, the purpose of the present invention is to provide a bioinformatics analysis tool that utilizes comparative genomics and recombinase Design of specific primers, probes and kits for the detection of Pythium oligandrum by polymerase amplification

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
  • Primer probe composition for detecting pythium oligandrum, kit, application and detection method
  • Primer probe composition for detecting pythium oligandrum, kit, application and detection method
  • Primer probe composition for detecting pythium oligandrum, kit, application and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: Design, screening and verification of specific primers and probe compositions of Pythium oligandrum

[0042] In order to obtain the specific primer and probe composition of Pythium oligandrum, first, based on bioinformatics tools, using comparative genomics methods, sequence-specific genes were identified on the Pythium oligandrum genome as detection targets. First, the nucleic acid sequences of all the genes on the Pythium oligandrum genome were compared to the other nine sequenced Pythium species (Pythium irregulare, Pythium insidiosum, Pythium vexans, Pythium myriotylum, Pythium brassicum, Pythium ultimum, Pythium arrhenomanes, Pythium guiyangense, Pythium iwayamai) all gene sequences, and found 189 Pythium malogyno genes that have no homologous genes in other Pythium species. Then, by comparing with NCBI's NR database Blast, 6 relatively specific genes were selected as detection targets, and 6 sets of primer compositions were designed.

[0043] Primer...

Embodiment 2

[0062] Embodiment 2: the specificity test of the primer of Pythium oligandrum and probe composition

[0063] Primer and probe combination for detecting Pythium oligandrum: the forward primer oilF has the nucleotide sequence shown in SEQ ID No.1, and the reverse primer B-oilR has the nucleotide sequence shown in SEQ ID No. The nucleotide sequence shown in 2, the probe T-oil has the nucleotide sequence shown in SEQ ID No.3.

[0064] The above-mentioned primer-probe composition was used to prepare a kit, and the concentration and dosage of each reagent in the kit were: 2.1 μL 10 μM forward primer oilF, 2.1 μL 10 μM reverse primer B-oilR, 0.6 μL 10 μM probe T-oil, 29.5μL Rehydration Buffer, 11.2μL DEPC-treated water, 2.5μL 280mM MgAC and dry enzyme powder were used to prepare a 48μL reaction system.

[0065] Extract the genomic DNA of the sample to be tested, take 2 μL of the sample DNA as a reaction template, add 48 μL of the detection solution in the kit to carry out recombinas...

Embodiment 3

[0067] Embodiment 3: the sensitivity test of Pythium oligandrum recombinase polymerase detection

[0068] In order to determine the sensitivity of the recombinant enzyme polymerase detection method, the genomic DNA of Pythium oligandrum was measured with a spectrophotometer and then diluted 10 times. The DNA concentration range was set at 10ng-10fg, and the DNA of each concentration after dilution was diluted. 2 μL of solution was used as a template, and DEPC-treated water was used as a negative control, and 48 μL of the kit solution was added to carry out the recombinant enzyme polymerase amplification reaction. The reaction procedure was: incubate at 39°C for 25 minutes. After the reaction, use lateral flow chromatography test strips to detect the amplification product, and judge the detection result according to the color change of the detection line indicator band. The result shows: DNA concentration can observe band at test strip detection line place in the scope of 10ng-...

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 primer probe composition for detecting pythium oligandrum, a kit, application and a detection method, and relates to the technical field of gene engineering. A primer probe composition identified on a pythium oligandrum genome by utilizing a comparative genomics method consists of a forward primer oilF, a reverse primer B-oilR and a probe T-oil. Compared with a traditional detection technology for identifying pythium oligandrum according to morphological characteristics, the method has higher accuracy, sensitivity and effectiveness, is convenient to operate and good in practicability, provides a new technical platform for detection of pythium oligandrum, and can be used for high-sensitivity rapid detection and screening of biocontrol bacteria with antagonism and growth promotion effects.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a primer probe composition, a kit, an application and a detection method for detecting Pythium oligandrum. Background technique [0002] Soil-borne pathogens are an important factor causing crop yield reduction. Although traditional measures such as changing farming patterns can inhibit the spread of soil-borne pathogens to a certain extent, due to the characteristics of long-term prevalence and rapid onset of pathogens, plant diseases still cause global agricultural economy. immeasurable loss. Although chemical pesticides can effectively inhibit the growth of pathogenic bacteria, excessive use or abuse has caused serious harm to the environment and human health. Therefore, the use of environmentally friendly plant probiotics to antagonize soil-borne pathogenic bacteria has become a research hotspot in recent years. Pythium oligandrum belongs to the funga...

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 Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6844C12N15/11C12R1/645
CPCC12Q1/6895C12Q1/6844C12Q2521/507C12Q2522/101C12Q2531/119C12Q2537/1376C12Q2565/625Y02A50/30
Inventor 柴春月惠丰立刘金丽陈崇艺朱露露
Owner NANYANG NORMAL 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