Kit for detecting novel coronaviruses (2019-nCoV) based on double amplification technology and application of kit

A technology for 2019-ncov and coronavirus, applied in the detection/inspection of microorganisms, resistance to vector-borne diseases, biochemical equipment and methods, etc., can solve the problems of unfavorable popularization and application, and achieve the effect of improving detection efficiency and sensitivity

Active Publication Date: 2020-06-26
武汉中帜生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the PCR method has certain requirements on hardware facilities, and requires a dedicated PCR diagnostic laboratory and expensive experimental equipment, which is not conducive to popularization and application in some communities and remote hospitals.

Method used

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  • Kit for detecting novel coronaviruses (2019-nCoV) based on double amplification technology and application of kit
  • Kit for detecting novel coronaviruses (2019-nCoV) based on double amplification technology and application of kit
  • Kit for detecting novel coronaviruses (2019-nCoV) based on double amplification technology and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] [Example 1] Sensitivity Test

[0086] Select a known concentration of the pseudovirus containing the 2019-nCoV target gene, perform a 10-fold concentration gradient dilution, and repeat each gradient dilution three times separately, and use the lowest dilution concentration with a 100% positive detection rate as the estimated detection limit , after the estimated detection limit is determined, dilute the 2019-nCoV pseudovirus to near the estimated detection limit concentration, and use the kit for detection, each concentration is tested 20 times to further accurately determine the minimum detection limit concentration (the positive rate is selected at The dilution above 95% is used as the detection limit sensitivity of this kit).

[0087] Table 1 2019-nCoV Pseudovirus Experiment Results - Determination of Estimated Detection Limit

[0088]

[0089] Table 2 Experimental results of 2019-nCoV pseudovirus - determination of detection limit

[0090]

[0091]

[00...

Embodiment 2

[0093] [Example 2] Specificity Verification

[0094] Other pathogens that are similar to the 2019 novel coronavirus species or cause similar symptoms (such as seasonal influenza A H1N1 virus, novel influenza A H1N1 (2009) virus, influenza A H3N2, H5N1, H7N9, influenza B Yamagata, influenza B Influenza type Victoria, RSV type A, RSV type B, parainfluenza type I, parainfluenza type II, parainfluenza type III, rhinovirus group A, B, C, adenovirus 1, 2, 3, 4, Types 5, 7, 55, enterovirus A, B, C, D, human metapneumovirus (human metapneumovirus), Epstein-Barr virus, measles virus, human cytomegalovirus, rotavirus, norovirus, Mumps virus, varicella-zoster virus, Mycoplasma pneumoniae, Chlamydia pneumoniae, Legionella, Bordetella pertussis, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes, Klebsiella pneumoniae, tuberculosis Mycobacterium, Aspergillus fumigatus, Candida albicans, Candida glabrata, Cryptococcus neoformans, coronavirus (HK...

Embodiment 3

[0099] [Example 3] Validation of clinical samples

[0100] 1, reagent of the present invention and reference reagent detection result analysis

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PUM

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Abstract

The invention discloses a kit for detecting novel coronaviruses (2019-nCoV) based on a double amplification technology and an application of the kit. A collected sample is lysed by a cell lysis solution and then pathogen nucleic acid is released; and amplification of a pathogen nucleic acid fragment is realized through reverse transcription and transcription processes. An amplified RNA product isadded to a micropore coated with a coating probe, and a specific probe and an amplification probe are added at the same time, wherein the coating probe can be combined with one end of the specific probe CES to fix the amplified RNA product; and one end of the specific probe LES is bonded to the RNA product, and the other end of the specific probe LES is combined with the amplification probe to realize signal amplification. RNA extraction is not needed, and the kit is not easy to pollute in detection, has high sensitivity and strong specificity, can quickly and efficiently detect the novel coronaviruses (2019-nCoV), and has great help for epidemic spread and prevention.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a kit for detecting novel coronavirus (2019-nCoV) nucleic acid based on double amplification (RNA constant temperature amplification + multi-biotin signal amplification) technology and its application. Background technique [0002] Coronaviruses are non-segmented, single-stranded positive-sense RNA viruses belonging to the order Nidovirales, Coronaviridae, and Orthocoronavirinae, which are divided into subfamilies according to serotype and genomic characteristics. α, β, γ, and δ genera. 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 (MERSr-CoV) and severe acute respiratory syndrome-associated coronavirus (SARSr-CoV). CoV). The novel coronavirus 2019-nCoV is a novel coronavirus belonging to the genus Beta. At present, there is relatively ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/682C12Q1/6834
CPCC12Q1/682C12Q1/6834C12Q1/701C12Q2600/166C12Q2531/119C12Q2521/107C12Q2545/101Y02A50/30
Inventor 李先强姜昕黄永伟陈巨周向珍
Owner 武汉中帜生物科技股份有限公司
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