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Nucleic acid detection method for 2019 novel coronavirus

A coronavirus and detection method technology, applied in the field of 2019 novel coronavirus nucleic acid detection kits, can solve the problem of no specific treatment for the disease, and achieve the effects of short detection cycle, strong specificity and high sensitivity

Inactive Publication Date: 2020-05-29
拜澳泰克(沈阳)生物医学集团有限公司
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Problems solved by technology

There is currently no specific treatment for the disease caused by the new coronavirus

Method used

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  • Nucleic acid detection method for 2019 novel coronavirus
  • Nucleic acid detection method for 2019 novel coronavirus
  • Nucleic acid detection method for 2019 novel coronavirus

Examples

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example 1

[0028] Example 1: Use of the 2019 Novel Coronavirus Gene Detection Kit.

[0029] (1) Design of primers for the conserved region ORF1ab and Surface glycoprotein genes of the 2019 novel coronavirus genome:

[0030] ORF1ab-F: 5'-ACCTCATGGTCATGTTATGG-3';

[0031] ORF1ab-R: 5'-GACATAGCGAGTGTATGCC-3';

[0032] Surface glycoprotein-F: 5'-TTGGCAAAATTCAAGACTCACTTT-3';

[0033] Surface glycoprotein-R: 5'-TGTGGTTCATAAAAATTCCTTTGTG-3'.

[0034] (2) RNA extraction and amplification

[0035] 1. cDNA extraction: Collect samples and use RNA extraction solution to extract sample RNA and reverse transcribe it into cDNA.

[0036] 2. Take the reagents out of the refrigerator at -20°C first, put solution C on ice or at 4°C to melt the reagents, other reagents can be melted at room temperature.

[0037] 3. Add the following reagents to the 200μl PCR tube: the specific ratio is according to figure 1 join in.

[0038] 4. Use a 20μl volume pipette to gently pipette up and down to mix. Centrifu...

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Abstract

The invention provides a detection method and detection reagent kit for detecting 2019 novel coronavirus. PCR primers are two pairs of primers designed for detection of novel coronavirus ORF1ab and Surface glycoprotein genes. The detection reagent kit comprises Taq DNA polemerase mixed liquid, mixed liquid of the two pairs of primers, ddH2O, Marker, positive controls and negative controls. The detection method comprises the steps of using obtained sample cDNA as a template, and performing a PCR reaction by using the two pairs of primers; and performing agarose gel electrophoretic analysis on reaction products. According to the technical scheme provided by the invention, the 2019 novel coronavirus ORF1ab and Surface glycoprotein genes can be effectively detected. The method is high in specificity, high in sensitivity, short in detection period, rigorous and reliable in results and high in practicality. The invention provides an effective method for nucleic acid detection of the 2019 novel coronavirus.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnosis, and in particular relates to a nucleic acid detection kit and detection method for 2019 novel coronavirus. Background technique [0002] Coronaviruses are large, enveloped, positive-strand RNA viruses belonging to the Posterior Coronaviridae subfamily of the Coronaviridae family of the order Nidoviridae, which can be divided into 4 genera: α, β, δ, γ, which can cause colds and Middle East respiratory syndrome Severe acute respiratory syndrome (SARS) and severe acute respiratory syndrome (SARS). The novel coronavirus COVID-19, which has been infected since December 2019, belongs to the genus Betacoronavirus. The World Health Organization (WHO) named the pathogenic virus COVID-19 on February 11. The clinical manifestations of people infected with coronavirus are fever, cough, shortness of breath and dyspnea. In more severe cases, infection can lead to pneumonia, severe acute respirato...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/701C12Q1/686
Inventor 尹秀山尹瀛浩单国峰
Owner 拜澳泰克(沈阳)生物医学集团有限公司
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