RPA-LFD kit for rapidly detecting novel coronavirus

A coronavirus and kit technology, applied in the field of RPA-LFD kits, to achieve the effect of enhancing accuracy, reducing the burden on medical care, and shortening the detection time

Pending Publication Date: 2022-04-29
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, the adaptive evolution of the virus makes the task of finding the source of infection and cutting off the transmission route more difficult, and diagnostic tools must ensure that new mutant strains are still detectable
In addition, the currently used SARA-CoV-2 detection method is mainly fluorescent quantitative PCR, which requires professional equipment and technical personnel. Multiple large-scale screenings in a short period of time have brought a huge burden to the medical industry and front-line medical staff

Method used

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  • RPA-LFD kit for rapidly detecting novel coronavirus
  • RPA-LFD kit for rapidly detecting novel coronavirus
  • RPA-LFD kit for rapidly detecting novel coronavirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1 is used for the establishment of the RPA-LFD kit and method for rapid detection of novel coronavirus

[0062] 1. Materials and methods

[0063] 1.1 Virus strains and nucleic acids

[0064]The N and M gene fragments of the pcDNA3.1 vector pSARS-CoV-2 were connected. Avian influenza (AIV) H3N8, H5N6, H5N8, H7N9 and H9N2 subtypes, Newcastle disease virus (NDV), avian infectious bronchitis virus (IBV), parainfluenza B Virus (Sendaivirus, SeV), porcine epidemic diarrhea virus (porcine epidemic diarrhea virus, PEDV), porcine enteric alphacoronavirus (porcine enteric alphacoron virus, PEAV)

[0065] 1.2 Main instruments

[0066] BIOBASE biological safety cabinet, BIO-RAD gradient PCR amplification instrument, BIO-RAD fluorescence quantitative PCR amplification instrument, NanoDrop spectrophotometer, vortex instrument, instant centrifuge, Eppendorf centrifuge, BIO-RAD multi-purpose electrophoresis instrument.

[0067] 1.3 Main reagents

[0068] DH-5α from Heal...

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Abstract

The invention provides a composition, an RPA-LFD kit and a method for rapidly detecting novel coronavirus. The composition, the RPA-LFD kit and the method have the following advantages: (1) the detection time is greatly shortened, the large-scale screening speed can be effectively improved, and the medical burden is relieved; and (2) double-gene target detection: the accuracy of SARS-CoV-2 detection is enhanced. And (3) convenience: the RPA reaction is carried out under a constant temperature condition, expensive professional instrument support is not needed, the LFD detection method is simple and easy to operate, professional service is not needed, and on-site rapid diagnosis can be realized. (4) the specificity is good, namely the primer can be distinguished from other animal coronaviruses and also can be distinguished from various subtype influenza viruses, and the specificity is relatively good.

Description

technical field [0001] The invention relates to a RPA-LFD kit for rapid detection of novel coronavirus. Background technique [0002] Lambda and Mu are considered to be variants of interest (VOI) with predictable virulence enhancement, diagnostic escape or immune escape, and are being further monitored. my country has included COVID-19 as a Class B infectious disease stipulated in the "Law of the People's Republic of China on the Prevention and Control of Infectious Diseases". The prevention and control measures adopted for it are the same as those of Class A epidemic diseases. [0003] The full length of the SARS-CoV-2 genome is 29.9-32kb, consisting of 11 open reading frames (Open reading frame, ORF), 4 structural protein genes and an essential protein RNA-dependent RNA polymerase (RNA dependent RNA polymerase, RdRp) , Structural proteins include encoding spike glycoprotein S (Spike), envelope protein E (Envelope), membrane protein M (Membrane) and nucleoprotein N (Nuclec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6844C12Q2521/507C12Q2522/101C12Q2531/119C12Q2563/107C12Q2565/625C12Q2563/131
Inventor 任涛蔡琳琳向斌梁健鹏林秋燕陈礼斌
Owner SOUTH CHINA AGRI UNIV
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