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

Primer pair, probe and kit for detecting novel coronavirus SARS-CoV-2 and application of kit

A coronavirus, sars-cov-2 technology, applied in the direction of DNA / RNA fragments, microorganisms, recombinant DNA technology, etc., can solve the problems of backlog of suspected case samples, inability to get results quickly, inability to apply detection, etc., to achieve strong specificity , Overcoming the effects of long detection time and high sensitivity

Pending Publication Date: 2021-01-19
SHANGHAI BIOMEDICAL LAB CO LTD
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the RT-PCR method cannot be applied to basic level and on-site detection due to its temperature-variable characteristics that require related expensive and complex instruments and equipment, and requires professional laboratories and technicians.
Moreover, according to different kits, the RT-PCR method takes 1.5-3 hours for the entire detection time. Limited by the detection speed, it will lead to a backlog of samples of a large number of suspected cases, and the results cannot be obtained quickly.

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 pair, probe and kit for detecting novel coronavirus SARS-CoV-2 and application of kit
  • Primer pair, probe and kit for detecting novel coronavirus SARS-CoV-2 and application of kit
  • Primer pair, probe and kit for detecting novel coronavirus SARS-CoV-2 and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Design of primers and probes for detection of novel coronavirus SARS-CoV-2 and RPA detection method

[0026] 1. Design of primers and probes

[0027] Through comprehensive analysis and comparison of the published full sequence of the new coronavirus SARS-CoV-2 including 10 genes, the N and S gene sequences were selected for primer probe design. Through screening, two sets of primer probe sets listed in Table 1 below are preferably selected, wherein the first set is the upstream primer shown in SEQ ID NO.1 and the downstream primer shown in SEQ ID NO.3 designed for the N gene sequence , and the probe shown in SEQ ID NO.5; the second group is the upstream primer shown in SEQ ID NO.2, the downstream primer shown in SEQ ID NO.4, and the downstream primer shown in SEQ ID NO.6 designed for the S gene sequence probe.

[0028] Table 1 Sequences of primers and probes for the detection of novel coronavirus SARS-CoV-2

[0029]

[0030] Preferably, the 29th base of ...

Embodiment 2

[0038] Example 2 Detection method of rt-RPA for novel coronavirus SARS-CoV-2

[0039] 1) Take out the required components of the TAEXO02KIT kit 30 minutes in advance, thaw at room temperature, and mix by shaking.

[0040] 2) The primers and probes were dissolved in water, and the final concentration was 10 μM.

[0041] 3) Add 29.4 μL A buffer, 3 μL upstream primer, 3 μL downstream primer, 2 μL probe, 5.1 μL RNase free H to each dry powder reaction tube 2 O, invert the reaction tube 8-10 times and mix thoroughly.

[0042] 4) Add 5 μL RNA template and 2.5 μL B buffer to the reaction tube, invert up and down 8-10 times to mix, then centrifuge the reaction solution to the bottom of the tube briefly, and then immediately put the reaction tube into the fluorescence detection equipment (such as constant temperature expansion) increaser, real-time quantitative PCR, etc.).

[0043] 5) The constant temperature is 37°C; the fluorescence signal is collected every 30s (the selection of ...

Embodiment 3

[0045] Example 3 Optimization of Primer Concentration

[0046] with 10 3 Copie / μL plasmid standard is used to optimize the primer concentration, and the specific implementation is as follows:

[0047] 1) Take out the required components of the TAEXO02KIT kit 30 minutes in advance, thaw at room temperature, and mix by shaking.

[0048] 2) The primers and probes were dissolved in water, and the final concentration was 10 μM.

[0049] 3) Set up four experimental tubes, add 29.4 μL A buffer and 2 μL probe to each dry powder reaction tube, add 2 μL upstream primer and 2 μL downstream primer to 0.4 μM group, and add 3 μL upstream primer and 3 μL downstream primer to 0.6 μM group, 1.0 5μL upstream primer and 5μL downstream primer were added to μM group, NC group was a negative control, and an appropriate amount of RNase free H was used. 2 O Make up the system to 42.5 μL, invert the reaction tube up and down 8-10 times, and mix well.

[0050] 4) Add 5 μL RNA template and 2.5 μL B bu...

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 pair, probe and kit for detecting a novel coronavirus SARS-CoV-2. The kit comprises an upstream primer SEQ ID NO.1 and a downstream primer SEQ ID NO.3, or an upstreamprimer SEQ ID NO.2 and a downstream primer SEQ ID NO.4. The kit for detecting the novel coronavirus SARS-CoV-2 carries out nucleic acid detection on the SARS-CoV-2 by an RPA method, has the sensitivity of 250 copies / mL and the detection accuracy of 100%, can complete the detection of the novel coronavirus SARS-CoV-2 within 30 min, is very suitable for on-site rapid screening and has very good application prospects.

Description

technical field [0001] The present invention relates to the technical field of virus detection, in particular to primer pairs, probes, kits and applications for detecting novel coronavirus SARS-CoV-2. Background technique [0002] Coronavirus (Coronavirus) virus is a single-stranded positive-stranded RNA virus with an envelope (envelope), with a diameter of about 80-120 nm, its genetic material is the largest among all RNA viruses, and is an important pathogen of many domestic animals, pets, including human diseases. , can cause a variety of acute and chronic diseases. The Ninth Report of the International Committee on Taxonomy of Virology divides the Coronaviridae into three genera, namely alpha, beta and gamma. Among them, five kinds of viruses, including human coronavirus 229E of alpha coronavirus, human coronavirus NL63 and human coronavirus HKU1 of beta coronavirus, human coronavirus OC43, and severe acute respiratory syndrome (SARS) related virus, can be cause variou...

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): C12Q1/70C12N15/11C12R1/93
CPCC12Q1/701Y02A50/30
Inventor 周寅杜晓利杨紫瑶
Owner SHANGHAI BIOMEDICAL LAB 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