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

Method and kit for detecting novel coronavirus and D614G mutant strain thereof

A D614G, coronavirus technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial assay/inspection, etc., can solve problems such as unfavorable virus discovery and prevention, missed detection of mutant virus, etc.

Pending Publication Date: 2021-10-08
DAAN GENE CO LTD
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in view of the rapid mutation rate of RNA viruses, it is easy to cause missed detection of mutant viruses, which is not conducive to the timely detection and prevention of viruses

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
  • Method and kit for detecting novel coronavirus and D614G mutant strain thereof
  • Method and kit for detecting novel coronavirus and D614G mutant strain thereof
  • Method and kit for detecting novel coronavirus and D614G mutant strain thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Embodiment 1 kit and detection method

[0068] Design of primers and probes: Analyze the conserved regions of the genome based on the sequence of the novel coronavirus. Specific primers and probe sequences for the detection of the D614G mutation in the N gene and S protein of the 2019 novel coronavirus were designed in these conserved regions.

[0069] In addition, in order to monitor the process of sample collection, nucleic acid extraction and PCR amplification, ribonuclease P (RPP30) in human genomic DNA was selected to design internal standard primers and probes.

[0070] During the design process of the above primers and probes, try to avoid the formation of hairpin structures, internal dimers of primers, dimers between primers and mismatches. In addition, the above-mentioned novel coronavirus-specific primers and probe sequences were compared and analyzed by the NCBI Blast online database (https: / / blast.ncbi.nlm.nih.gov / Blast.cgi) to avoid comparison with other v...

Embodiment 2

[0087] The detection of embodiment 2 sensitivity

[0088] Connect the target fragment into the constructed pET28a-MS2 vector and transform the expression host strain BL21 competent cells, pick a single clone and sequence it for verification, induce expression, and digest the completely fragmented expression product with RNaseA and DNaseⅠ. Virus-like particles containing fragments of interest. Dilute the pseudovirus after the concentration determination to the appropriate concentration and then perform 10-fold dilution, the concentrations are 1.00E+07, 1.00E+06, 1.00E+05, 1.00E+04, 1.00E+03copies / ml. With the above-mentioned determined detection system and cycle parameters, the above-mentioned pseudovirus is detected ( Figure 5 ). The results show that the kit of the present invention has high sensitivity and can detect samples of 1.00E+03copies / ml.

Embodiment 3

[0089] Embodiment 3 specificity, repeatability detection

[0090]Influenza A / B virus, coronavirus 229E, coronavirus NL63, coronavirus OC43, coronavirus HKU1, respiratory syncytial virus, adenovirus, mycoplasma pneumoniae, Epstein-Barr virus, and chlamydia pneumoniae were tested as specific samples ( Figure 6 ). The results showed that this detection method has high specificity.

[0091] Selecting the pseudovirus that concentration is 1.00E+06copies / ml and 1.00E+04copies / ml detects, each repeats 10 times ( Figure 7 ). The results showed good repeatability.

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 method and a kit for detecting the novel coronavirus and a D614G mutant strain thereof, and particularly, through multiple rounds of screening and verifying, a primer probe set which is high in sensitivity, high in specificity, good in repeatability, high in anti-interference capability and capable of carrying out multiple detection is obtained from a large number of primer probe sets, and the primer probe set can be used for detecting the novel coronavirus and identifying a D614G mutant strain.

Description

technical field [0001] The invention belongs to the field of biotechnology and molecular diagnosis, in particular, the invention relates to a real-time fluorescent RT-PCR dual detection system for a novel coronavirus and its D614G mutant strain. Background technique [0002] The novel coronavirus is a single-stranded positive-sense RNA virus belonging to the family Coronaviridae and the subgenus Sarbecovirus in the genus Betacoronavirus. Because the new coronavirus is a single-stranded RNA virus, it is prone to mutations. Virus mutation and recombination are closely related to virus infectivity, reproduction ability, virus detection, and antibody effectiveness. Therefore, the monitoring of virus mutation and recombination is a crucial part of virus research. The S protein is one of the main proteins of the virus, which infects the host by interacting with the host receptor (ACE2), and determines the infecting target and infecting ability of the virus. [0003] The only sig...

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): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2600/16C12Q2600/166C12Q2521/107C12Q2531/113C12Q2537/143C12Q2563/107C12Q2545/113Y02A50/30
Inventor 蒋析文范建廖芷卉杨孟霞
Owner DAAN GENE CO LTD
Features
  • Generate Ideas
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com