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

Loop-mediated isothermal amplification detection primer of Pheretima cantonensis and method for identifying Pheretima cantonensis based on LAMP technology

A loop-mediated isothermal, primer-detecting technology, applied in recombinant DNA technology, biochemical equipment and methods, and microbial assay/inspection, etc., to achieve the effect of simple operation and low cost

Active Publication Date: 2019-01-11
INST OF ZOOLOGY GUANGDONG ACAD OF SCI
View PDF6 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the identification of earthworm species using LAMP technology

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
  • Loop-mediated isothermal amplification detection primer of Pheretima cantonensis and method for identifying Pheretima cantonensis based on LAMP technology
  • Loop-mediated isothermal amplification detection primer of Pheretima cantonensis and method for identifying Pheretima cantonensis based on LAMP technology
  • Loop-mediated isothermal amplification detection primer of Pheretima cantonensis and method for identifying Pheretima cantonensis based on LAMP technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] 1. Extraction of DNA

[0035] Take 30 mg of fresh and alive Cantonisosaurus (Pheniosaurus chinensis) tissue, and use an animal tissue DNA extraction kit (Tiangen) to extract DNA to obtain the DNA template of Cantonisosaurus.

[0036] 2. Primer design for Cantonosaurus

[0037] Through sequencing and calibration, the COI gene sequence of Guangdilong was obtained (as shown in SEQ ID NO.5), and with the COI gene as the target gene sequence, 4 primers were designed online using Primer Explorer V5software, and the obtained primer sequences were as follows:

[0038] Upstream inner primer FIP: 5'-CAACAGCCGCAGACCTTACTA-TTTT-AACATAAGATTTTGACTTTTGCC-3' (as shown in SEQ ID NO.1);

[0039] Downstream inner primer BIP: 5'-GGACAGTTTACCCCCCTTTAGC-TTTT-GCTAAATGTAGTGAGAAAATTGCA-3' (as shown in SEQ ID NO.2);

[0040] Upstream outer primer F3: 5'-CATAGCATTCCCACGTCTA-3' (as shown in SEQ ID NO.3);

[0041] Downstream outer primer B3: 5'-ACCTAAAATTGATGAGGCAC-3' (as shown in SEQ ID NO.4).

...

Embodiment 2

[0047] Sensitivity detection: the DNA template of Guangdilong extracted in Example 1 was divided into 1, 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 The dilution gradient was diluted, and LAMP (method was the same as in Example 1) and PCR were carried out respectively, and the sensitivity of the two identification methods was compared. Test results( image 3 ) shows that the sensitivity of LAMP method is 1000 times that of conventional PCR.

Embodiment 3

[0049] A total of 12 samples were collected by collecting live Earthworm from different places, dry products purchased from Chinese herbal medicine markets, and dried products purchased online. the result shows( Figure 4 , Figure 5 ) Samples 1, 7, 8, and 12 are Guangdilong, and the rest are other varieties. It was shown that the detection primers of the loop-mediated isothermal amplification of Guangdilong had good specificity.

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 loop-mediated isothermal amplification detection primer of Pheretima cantonensis and a method for identifying Pheretima cantonensis based on an LAMP technology. The inventiondesigns and screens a set of specific detection primers, a detection kit containing the detection primers and an identification method for determining whether a sample to be tested is Cantonese earthworm through loop-mediated isothermal amplification by using the detection kit for determining whether the sample is Cantonese earthworm or not. The invention is simple in operation, the result detection only needs naked eye observation, does not need expensive detection instrument, and the sensitivity is 1000 times higher than that of conventional PCR. The invention has lower cost and can be applied to authenticity identification of earthworm commodities.

Description

Technical field: [0001] The invention belongs to the technical field of resource identification of traditional Chinese medicinal materials, and in particular relates to a detection primer for loop-mediated isothermal amplification of A. guangtilong and a method for identifying A. guangtilong based on LAMP technology. Background technique: [0002] Guangdilong is an annelid phylum Megalucidae. It is a traditional animal Chinese medicinal material with a long history of medicine. Guangdilong and its preparations are widely used in clinical practice. Guangdilong has a variety of pharmacological effects, mainly including lowering blood pressure, relieving asthma, antipyretic and anti-inflammatory, antithrombotic, and antitumor. Dilong is a traditional folk medicine in China. According to the Chinese Pharmacopoeia, Dilong includes Guangdilong and Hudilong. The quality of Dilong is considered to be the best in the industry. There are many varieties in China, and the shape and siz...

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): C12Q1/6888C12Q1/6844C12N15/11
CPCC12Q1/6844C12Q1/6888C12Q2531/119C12Q2565/125C12Q2563/107
Inventor 黄庆毛润乾李志武
Owner INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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