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Novel coronavirus integrated nucleic acid rapid detection card box

Technology of a coronavirus, test card, used in the field of medical testing

Active Publication Date: 2021-05-07
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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  • Novel coronavirus integrated nucleic acid rapid detection card box
  • Novel coronavirus integrated nucleic acid rapid detection card box
  • Novel coronavirus integrated nucleic acid rapid detection card box

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 novel 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 PCR tube. And add 5ul template RNA.

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

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

Embodiment 2

[0064] Example 2. Creation of reagent cartridges.

[0065] 1. The reagent cartridge includes the following components:

[0066] ①Primer probe combination

[0067]

[0068] ②Detection solution components:

[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 concentration of the probe used to detect the S target gene is 200nM, the concentration of the probe used to detect the N target gene is 200nM, the concentration of the probe used to detect the internal standard is 100nM, used for The concentration of upstream and downstream primers used to detect the S target gene is 400nM, the concentration of upstream and downstream primers used to detect the N target gene is 400nM, and the concentration of upstream and downstream primers used to detect the internal standard is 200nM;

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

[0071] ③ Purifi...

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 the detection of 2019-nCoV, we tested the sensitivity and specificity of the cartridge.

[0101] 1.1 Test the 2019-nCoV quality control products at different concentrations 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, and 3, Adenovirus, rubella virus, vesicular stomatitis virus, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pneumoniae, Mycobacterium tuberculosis, Candida albicans, Candida glabrata and human genome were tested to verify specificity

[0103] 2. Results:

[0104] 2.1 We tested different concentrations of 2019-nCoV quality control products, and the following 2 tables are the results of S gene and N gene respectively.

[010...

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Abstract

The invention discloses a novel coronavirus integrated nucleic acid rapid detection card box. The novel coronavirus integrated nucleic acid rapid detection card box comprises a box body with a box cover at the top, wherein a cracking cavity with a cracking liquid, a first cleaning cavity with a first cleaning liquid and a second cleaning cavity with a second cleaning liquid are sequentially arranged in the box body from top to bottom, the cracking cavity communicates with the box cover, and the second cleaning cavity communicates with a reaction cavity arranged at the bottom of the outer side of the box body; plungers with plunger hole directions perpendicular to the connecting direction are respectively arranged in the connecting sections of every two of the cracking cavity, the first cleaning cavity, the second cleaning cavity and the reaction cavity, and hydrophobic liquid sealing materials are arranged in plunger holes of the plungers; an amplification reaction liquid and a novel coronavirus detection primer and probe combination are arranged in the reaction cavity; and magnetic beads capable of penetrating through the plunger holes of the plungers are arranged in the cracking cavity. The novel coronavirus integrated nucleic acid rapid detection card box disclosed by the invention can integrally perform nucleic acid extraction and PCR amplification on novel coronaviruses under the condition of relatively low cost, and further can directly perform data collection and analysis required by 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 as 2019-nCoV, and named the pneumonia caused by the virus as COVID-19. [0003] The current clinical application of novel coronavirus detection methods mainly include the following: [0004] (1) Isolation 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 for culture, and the cell lesions can be observed and the virus can be isolated to identify the virus. The cultivation period is long, and it needs to be carried out in a biosafety level 3 laboratory, so the application of this method in clinical diagnosis is limited. [0006] (2) Specific antibody detection [0007] After the virus infects t...

Claims

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Application Information

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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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