Real-time fluorescent PCR detection method for dendroctonus melanocarpa, primer and probe for detection

A detection method, a real-time fluorescence technology, is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., to achieve the effect of improving the detection rate, preventing invasion and proliferation, and intuitive identification results.

Pending Publication Date: 2021-06-25
海口海关热带植物隔离检疫中心
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no real-time fluorescent PCR rapid identification technology research for the molecular identification of beetles.

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 for dendroctonus melanocarpa, primer and probe for detection
  • Real-time fluorescent PCR detection method for dendroctonus melanocarpa, primer and probe for detection
  • Real-time fluorescent PCR detection method for dendroctonus melanocarpa, primer and probe for detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0038] Below, the technical solutions of the present invention will be further explained in conjunction with specific embodiments, but the present invention is not limited to the following embodiments.

[0039] A real-time fluorescent PCR detection method for the black fat beetle, specifically comprising the following steps:

[0040] (1) Design primers and probes

[0041] The following is the design and synthesis process of the specific primers and probes used in the detection method of the black fat beetle of the present invention.

[0042] Using MEGA6.0 software, the southern pine beetle D.frontalis, the yellow fir beetle D.pseudotsugae, the red beetle D.valens, the red wing beetle D.rufipennis, the Huashan pine beetle D.armandi, the black beetle beetle D.terebrans, D.adjunctus, D.approximatus, D.brevicomis, D.jeffreyi, D.micans, D. Murrayanae, D.ponderosae, D.punctatus, D.simplex, D.mexicanus, D.parallelocollis, D.rhizophagus mtCOI gene (mitochondrial DNA) sequences were...

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 provides a detection method for dendroctonus melanocarpa. The purpose of identifying dendroctonus melanocarpa can be achieved by combining specific primers (Heii-F and Heii-R) and a probe (Heii-P) which are screened by a test in a Taq-Man real-time fluorescent quantitative PCR method. The invention is suitable for quarantine identification of beetles in different states, the defects of a conventional morphological identification method are overcome, and compared with a DNA bar code identification technology applied in the prior art, the primer and the MGB probe are high in specificity and sensitivity, simple in experimental operation and short in consumed time; The identification result is visual and can be obtained by observing an amplification curve. The research not only can meet the original rapid customs clearance requirement of entry and exit, improve the detection rate, but also can effectively prevent invasion and diffusion of dangerous dendroctonus beetle, and has important significance on protection of agriculture and forestry production and ecological safety in China.

Description

technical field [0001] The invention belongs to the technical field of molecular identification of biological species, and relates to a real-time fluorescent PCR detection method for the beetle, in particular to a method for detecting the beetle and its application; in addition, the present invention also relates to primers and probes for detecting the beetle. Needle. Background technique [0002] The genus Dendroctonus belongs to Coleoptera, Curculionidae, Scolytinae, and is an important dry-boring pest in forest ecosystems. It mainly causes damage to trees such as Pinus, Picea, and Douglas. This genus (non-Chinese species) has been included in the list of imported plant quarantine pests in my country. There are a total of 20 known species of the genus Glycerus in the world, of which 18 species are distributed in Central and North America, and 3 species are currently distributed in Europe and Asia. Dendroctonus terebrans is a non-Chinese species of pest. The black beetle D...

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): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2561/113C12Q2563/107C12Q2545/114C12Q2561/101
Inventor 刘嘉欢孟瑞蔡波敖苏徐卫刘福秀韩玉春陈施明王安石
Owner 海口海关热带植物隔离检疫中心
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