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A novel coronavirus integrated nucleic acid rapid detection cartridge

A coronavirus and detection card technology, applied in the field of medical testing, to achieve the effect of eliminating aerosol pollution, protecting safety, and reducing the requirements for professional skills

Active Publication Date: 2022-08-02
杭州遂曾生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The vast majority of grassroots hospital laboratories do not have the above-mentioned professionals and equipment and facilities, and will only transport samples to qualified laboratories. In most cases, qualified laboratories cannot finish the samples of their own hospitals. The difficulty of diagnosis can be imagined

Method used

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  • A novel coronavirus integrated nucleic acid rapid detection cartridge
  • A novel coronavirus integrated nucleic acid rapid detection cartridge
  • A novel coronavirus integrated nucleic acid rapid detection cartridge

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1 Primer probe design and screening.

[0040] 1. The present invention analyzes and studies the sequence of the new coronavirus 2019-nCoV, and designs the primer probe sets shown in Table 1-Table 4.

[0041] Table 1 Primer probe of the present invention

[0042]

[0043]

[0044] Table 2 Control group 1 primer probe

[0045]

[0046] Table 3 Control group 2 primer probe

[0047]

[0048]

[0049] Table 4 Control group 3 primer probes

[0050]

[0051] 2. At the same time, through the optimization of the detection system, the following detection system was established: add 30ul RT-PCR amplification solution and 5ul manganese ion solution to each tube of PCR. And add 5ul template RNA.

[0052] 30ul RT-PCR amplification solution: including primers, probes and PCR buffer;

[0053] The probe used to detect the S target gene was used at a concentration of 200nM, the probe used to detect the N target gene was used at a concentration of 200nM, the...

Embodiment 2

[0064] Example 2. Creation of reagent cartridges.

[0065] 1. The kit includes the following components:

[0066] ①Primer probe combination

[0067]

[0068] ②Components of detection liquid:

[0069] The composition of PCR buffer is: 50mmol / L Tris-HCl (pH8.0), 20mmol / L (NH 4 )SO 4 , 30mmol / LKCl, 4.0mmol / L MgCl 2 , 0.2mmol / L dNTPs, 5U rTth DNA polymerase, the probe used to detect the S target gene was used at a concentration of 200nM, the probe used to detect the N target gene was used at a concentration of 200nM, and the probe used to detect the internal standard was used at a concentration of 100nM. The upstream and downstream primers used to detect the S target gene use a concentration of 400nM, the upstream and downstream primers used to detect the N target gene use a concentration of 400nM, and the upstream and downstream primers used to detect the internal standard use a concentration of 200nM;

[0070] 5ul manganese ion solution (25mM manganese acetate);

[007...

Embodiment 3

[0099] Example 3. Test Example 1.

[0100] 1. In order to test the accuracy and effectiveness of the cartridge of the present invention for 2019-nCoV detection, we tested the sensitivity and specificity of the cartridge.

[0101] 1.1 Detect different concentrations of 2019-nCoV quality control materials to confirm the detection limit;

[0102] 1.2 For endemic human coronaviruses (HKU1, OC43, NL63 and 229E), SARS coronavirus, MERS coronavirus, H1N1, H3N2, H7N9, influenza B, respiratory syncytial virus, parainfluenza virus types 1, 2, 3, Adenovirus, rubella virus, vesicular stomatitis virus, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pneumoniae, Mycobacterium tuberculosis, Candida albicans, Candida glabrata and human genome are tested to verify specificity

[0103] 2. Results:

[0104] 2.1 We tested different concentrations of 2019-nCoV quality control materials. The following two tables are the results of S gene and N gene respectively.

[0105]

[0106]...

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Abstract

The invention discloses a novel coronavirus integrated nucleic acid rapid detection cartridge, comprising a box body with a box cover on the top, and a lysis chamber with a lysis solution and a first cleaning solution with a first cleaning solution are arranged in the box body from top to bottom in order. a cleaning chamber and a second cleaning chamber with a second cleaning solution, wherein the cracking chamber is connected to the box cover, and the second cleaning chamber is connected to the reaction chamber provided at the bottom of the outer side of the box body; the cracking chamber, the first cleaning chamber, the second cleaning chamber The two connected sections of the cavity and the reaction cavity are respectively provided with plungers whose direction of the plunger hole is perpendicular to the connection direction, and the plunger hole of the plunger is provided with a hydrophobic liquid sealing material; the reaction cavity is provided with an amplification reaction solution , combined with novel coronavirus detection primers and probes; the lysis chamber is provided with magnetic beads that can pass through the plunger hole of the plunger. The present invention can perform nucleic acid extraction and PCR amplification for the novel coronavirus in an integrated manner at relatively low cost, and then directly perform data collection and analysis required for optical detection.

Description

technical field [0001] The invention relates to a novel coronavirus integrated nucleic acid rapid detection cartridge, which belongs to the technical field of medical detection. Background technique [0002] On January 12, 2020, the World Health Organization officially named the new coronavirus 2019-nCoV, and named the pneumonia caused by the virus as COVID-19. [0003] At present, the detection methods of new coronavirus in clinical application mainly include the following: [0004] (1) Isolation and culture [0005] The virus can infect a variety of human or non-human cell lines in vitro, and the virus can be inoculated on the cell line and cultured, and the cytopathic changes can be observed and the virus can be isolated, so as to identify the virus. The culture requires a long period of time and needs to be carried out in a biosafety tertiary laboratory. The application of this method in clinical diagnosis is limited. [0006] (2) Specific antibody detection [0007]...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/34C12M1/00C12Q1/70C12Q1/686
CPCC12Q1/701C12Q1/686C12Q2600/166C12Q2521/101C12Q2545/101C12Q2527/125C12Q2563/143C12Q2563/149Y02A50/30
Inventor 赵怀赵志敬华灿忠
Owner 杭州遂曾生物技术有限公司
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