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Novel coronavirus detection kit

A detection kit and coronavirus technology, applied in the field of new coronavirus detection kits, can solve problems such as difficulties in popularization and inability to perform visual interpretation of results, so as to relieve work pressure, reduce the possibility of cross-infection in hospitals, and be fast, accurate and convenient Detection effect

Pending Publication Date: 2020-08-07
亚能生物技术(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, the existing CRISPR / cas12 detection system needs the help of a fluorescence detector, and the results cannot be interpreted visually, which brings difficulties to the popularization of this technology in grassroots inspection institutions

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0036] A novel coronavirus detection kit, including crRNA, Cas12 protein, RPA upstream and downstream primers, buffer system and single-stranded DNA reporter molecule;

[0037] The crRNA is a specific crRNA designed for the target gene, with a length of 17-25 nucleotides. There should be a PAM sequence near the 5' end of the crRNA. The PAM sequence is TTTN, and the PAM sequence is used for crRNA and cas12 identification of the constituent complexes;

[0038] The crRNA and single-stranded DNA reporter molecules designed and chemically synthesized in this example are shown in Table 1 below.

[0039] Table 1

[0040]

[0041] The RPA upstream and downstream primers in this example are shown in Table 2 below.

[0042] Table 2

[0043] Oligo name Sequence(5'-3') 2019-nCoV-ORF1ab-F ATGGTGACTTTTTGCATTTCTTACCTAGAGTTT (SEQ ID NO: 1) 2019-nCoV-ORF1ab-R GCTGATGTTGCAAAGTCAGTGTACTCTATAAG (SEQ ID NO: 2) 2019-nCoV-N-F CAGGCAGCAGTAGGGGAACTTCTCCTGCTAGAAT ...

Embodiment 2

[0056] A novel coronavirus detection kit, including crRNA, Cas12 protein, RPA upstream and downstream primers, buffer system and single-stranded DNA reporter molecule;

[0057] The crRNA is a specific crRNA designed for the target gene, with a length of 17-25 nucleotides. There should be a PAM sequence near the 5' end of the crRNA. The PAM sequence is TTTN, and the PAM sequence is used for crRNA and cas12 identification of the constituent complexes;

[0058] The crRNA and single-stranded DNA reporter molecules designed and chemically synthesized in this example are shown in Table 6.

[0059] Table 6

[0060]

[0061] RPA upstream and downstream primers in this example are shown in Table 2.

[0062] The detection solution ratio of cas12a used in this example is shown in Table 7 below.

[0063] Table 7

[0064]

[0065] Use of the above kits:

[0066] 1) Use pseudoviral particles containing the genome sequence of the new crown, and use throat swabs to sample healthy v...

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Abstract

The invention relates to the technical field of gene detection, in particular to a novel coronavirus detection kit which comprises crRNA, Cas12 protein, RPA upstream and downstream primers, a buffer system and a single-stranded DNA reporter molecule. The crRNA is crRNA which is designed to be specific for a target gene; the upstream and downstream primers of the RPA are as follows: a 5'-TTTN-3' sequence is searched for aiming at a conserved region ORF1ab and an N gene region of the novel coronavirus, a forward primer is designed within 0-200 bp nearby the 5'-region of the novel coronavirus, and a reverse primer is designed within 25-200 bp nearby the near 3'-region of the novel coronavirus. The method has the advantages that due to the fact that RPA isothermal amplification has low requirements for samples, a rapid and simple sample treatment mode can be adopted, RPA-mediated DNA amplification can be conducted at the temperature of 30-42 DEG C, and dependence on a precise PCR instrument is avoided.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a novel coronavirus detection kit. Background technique [0002] Common signs after coronavirus infection are respiratory symptoms, fever, cough, shortness of breath, and dyspnea. In more severe cases, infection can lead to pneumonia, severe acute respiratory syndrome, kidney failure, and even death. However, due to relatively hidden factors such as the lack of obvious initial symptoms and the incubation period of up to 14 days, patients and carriers of the new coronavirus are often not detected in time, which intensifies the spread. [0003] Early and specific diagnosis of infected patients by rapid and effective means is an important means to detect and isolate the source of infection in time, effectively treat patients, and ensure social order. This puts forward new requirements for the rapid diagnosis technology of pathogens. However, when severe infectious disease...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6844C12Q2521/507C12Q2521/327C12Q2527/101
Inventor 汤光辉薛良代文俊李观得刘南松龚姣姣
Owner 亚能生物技术(深圳)有限公司
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