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

Composition, kit, method and application for detecting and typing coronavirus

A technology of coronaviruses and compositions, applied in the field of compositions, kits, and applications for detecting and typing coronaviruses, can solve problems such as prone to serious illness, and achieve avoidance of false positives and environmental pollution, high throughput, The effect of improving detection efficiency

Active Publication Date: 2020-05-22
SANSURE BIOTECH INC
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Among the seven coronaviruses that can infect humans, coronavirus 229E, coronavirus NL63, coronavirus OC43, and coronavirus HKU1 are common respiratory infection viruses. If infected by coronavirus 2019-nCoV, coronavirus MERSr-CoV, or coronavirus SARSr-CoV, the symptoms of the patient's infection will be very serious, and severe illness will easily occur.

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
  • Composition, kit, method and application for detecting and typing coronavirus
  • Composition, kit, method and application for detecting and typing coronavirus
  • Composition, kit, method and application for detecting and typing coronavirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Embodiment 1, primers and probes used in the present invention

[0052] The primers and probes used in the present invention are shown in Tables 1 to 3 below:

[0053]

[0054]

[0055]

[0056]

[0057]

[0058]

[0059] Among them, the fluorescent reporter group of the novel coronavirus 2019-nCoV probe is FAM; the fluorescent reporter group of the coronavirus NL63 probe is HEX; the fluorescent reporter group of the coronavirus HKU1 probe is ROX, and the 3' end of the probe is Also has a BHQ1 or BHQ2 quenching group.

[0060] The fluorescent reporter group of the upstream primer of coronavirus 229E is FAM; the fluorescent reporter group of the upstream primer of coronavirus OC43 is HEX; the fluorescent reporter group of the upstream primer of coronavirus MERSr-CoV is ROX; the fluorescent reporter group of coronavirus SARSr-CoV The group is CY5.

Embodiment 2

[0061] Embodiment 2, the method for detecting coronavirus and typing

[0062] The detection samples of the present invention are throat swabs, sputum, alveolar lavage fluid, and blood. Virus nucleic acid is extracted by the magnetic bead method, and the following operations are performed in the sample processing room:

[0063] 2.1 Take an appropriate amount of 1.5 mL sterilized centrifuge tubes, mark the negative control, positive control and samples to be tested respectively, and add 300 μL RNA extraction solution 1 to each tube;

[0064] 2.2 Add 200 μL of the sample to be tested or the negative control and positive control to each tube; cover the tube cap, shake and mix for 10 seconds, and centrifuge briefly;

[0065] 2.3 Add 100 μL RNA Extraction Solution 2-mix to each tube (mix well and draw), shake and mix for 10 seconds, then let stand at room temperature for 10 minutes;

[0066] 2.4 Place the centrifuge tube on the separator after brief centrifugation, and slowly suck...

Embodiment 3

[0082] Embodiment 3, the detection result of composition test positive control of the present invention

[0083] Using the compositions in Table 1 to Table 3 of the present invention, according to the method described in Example 2, each target-positive plasmid is detected to simulate clinical samples, and multiplex PCR detection is carried out on the Hongshi fluorescent quantitative PCR instrument. The results are as follows Figure 1~8 shown. It can be seen from the figure:

[0084] When the coronavirus 2019-nCoV is detected with the composition in Table 1, the Ct value is around 25; when it is detected with the composition in Table 2, the Ct value is around 28; when it is detected with the composition in Table 3, the Ct value is between Around 32, the amplification curves of A, B and C are relatively straight.

[0085] When coronavirus NL63 is detected with the composition in Table 1, the Ct value is around 30; when tested with the composition in Table 2, the Ct value is a...

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
diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to the field of molecular biology detection, and more particularly, to the field of detection of coronaviruses. The invention provides a composition capable of detecting novel coronavirus 2019-nCoV, coronavirus 229E, coronavirus NL63, and coronavirus OC43 , coronavirus HKU1, coronavirus MERSr-CoV, coronavirus SARSr-CoV and typing; at the same time, also provide a kit comprising the composition, the use of the composition, and for the detection of coronavirus and method of typing. The composition of the invention combines the fluorescent probe method and the melting curve method, and can simultaneously detect and type seven coronaviruses in one tube, with low cost, high throughput, less time-consuming, and greatly improved detection efficiency. And the operation is simple, and the single-tube operation avoids false positives and environmental pollution caused by crossover between samples.

Description

technical field [0001] The present invention belongs to the field of molecular biology detection, more specifically, belongs to the field of coronavirus detection, more specifically, the present invention can detect novel coronavirus 2019-nCoV, coronavirus 229E, coronavirus NL63, coronavirus OC43, coronavirus HKU1 , coronavirus MERSr-CoV, and coronavirus SARSr-CoV were typed. Background technique [0002] Coronaviruses are non-segmented, single-stranded, positive-sense RNA viruses that belong to the order Nidovirales, Coronaviridae, and Orthocoronavirinae. According to serotype and genomic characteristics, the coronavirus subfamily is divided into The four genera α, β, γ, and δ can infect many animal species, including humans, bats, dogs, pigs, mice, birds, cattle, whales, horses, goats, monkeys, etc. There are 6 coronaviruses known to infect humans, including 229E and NL63 of the genus α, OC43 and HKU1 of the genus β, Middle East respiratory syndrome-associated coronavirus...

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): C12Q1/70C12Q1/6851C12R1/93
CPCC12Q1/701C12Q1/6851C12Q2600/166C12Q2531/113C12Q2563/107C12Q2545/101C12Q2600/16C12Q1/6844C12Q2527/107C12Q2537/143C12Q2561/113C12Q2565/102
Inventor 戴立忠谭德勇邓中平纪博知刘佳程星孙青芝颜进毛君杰
Owner SANSURE BIOTECH INC
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