Novel coronavirus detection method and kit

A coronavirus and detection method technology, applied in the biological field, can solve problems such as lack of antiviral drugs, and achieve the effect of convenient preparation and pollution prevention

Pending Publication Date: 2020-06-26
SHANGHAI XINGYAO MED TECH DEV CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is a lack of effective antiviral drugs against patho

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Embodiment 1 novel coronavirus detection kit (fluorescent PCR method)

[0072] 1. Primer Probe Synthesis

[0073] Entrust Yingwei Jieji (Shanghai) Trading Co., Ltd. to synthesize the primer probe. The sequence of the synthesized primer probe is as follows:

[0074] The upstream primers are:

[0075] 5'- AATGATGATACTCTCTGACGATGCTGTTG -3' as shown in SEQ ID No.1;

[0076] 5'- GGAAGTCACACCTTCGGGAACGTGGTTG-3' as shown in SEQ ID No.4;

[0077] 5'- GTGTGCGTACTGCTGCAATATTGTTAACG-3' as shown in SEQ ID No.7;

[0078] Downstream primers are:

[0079] 5'-CATTGTTTTGATAATAAAGAACTGACTTAAAG-3', as shown in SEQ ID No.2;

[0080] 5'-CAGCAAAATGACTTGATCTTTGAAATTTGG-3', as shown in SEQ ID No.5;

[0081] 5'-ACCAGAAGATCAGGAACTCTAGAAG-3', as shown in SEQ ID No.8

[0082] The sequence of the novel coronavirus ORF1ab gene probe is:

[0083] 5'- GTTTCAATAGCACTTATGCATCTCAAGGTCTAGT-3', the 5' end is marked with FAM, and the 3' end is marked with BHQ1, as shown in SEQID No.3;

[0084] The ...

Embodiment 2

[0104] Application of embodiment 2 novel coronavirus detection kit (fluorescent PCR method)

[0105] 200 cases of clinical samples were tested by using the kit obtained in the present invention, and compared with the clinical diagnosis results to evaluate the effect of the kit.

[0106] 1. Detection of clinical samples with the new coronavirus detection kit (fluorescent PCR method)

[0107] (1) Reagent preparation

[0108] Prepare the reaction solution according to the number of reaction samples (number of reaction samples = number of samples to be tested + 2 reference substances + 1) n: take 2019-nCov reaction solution n×14 μl, RT-PCR enzyme n×6 μl and mix them in a centrifuge tube; Centrifuge at low speed for a few seconds, and dispense 20 μl / tube into reaction tubes. After dispensing, the reaction tube can be placed at 2-8°C for 3 hours.

[0109] (2) Nucleic acid extraction

[0110] Take 200 μL of the pharyngeal swab eluate or sputum digestion solution in the sampling t...

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PUM

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Abstract

The invention provides a novel coronavirus detection method and kit. The method adopts a fluorescent nucleic acid amplification method with multiple pairs of primers and multiple probes, and can realize detection of multiple target nucleic acid sequences in a single tube. The novel coronavirus ORF1ab gene, N gene and E gene are used as target sequences to design specific primers and TaqMan probes,and the novel coronavirus is detected through multi-channel fluorescent PCR amplification. The design of the primers and probes, a reaction system and an amplification program are optimized, the fluorescence detection can be completed in 70 minutes, and the sensitivity of the kit reaches 200 copies/mL.

Description

technical field [0001] The invention is a novel coronavirus detection method and kit, belonging to the field of biotechnology. Background technique [0002] Novel coronavirus pneumonia is an acute infectious pneumonia, and its pathogen is a new type of coronavirus that has not been found in humans before, that is, the 2019 novel coronavirus. On February 7, 2020, the National Health and Medical Commission decided to temporarily name "new coronavirus-infected pneumonia" as "new coronavirus pneumonia", or "new coronary pneumonia" for short. On February 11, the World Health Organization (WHO) named it Corona Virus Disease 2019 (COVD-19) in English. On February 22, the National Health and Medical Commission decided to revise the English name of "new coronavirus pneumonia" to "COVID-19", which is consistent with the naming of the World Health Organization, and the Chinese name remains unchanged. On January 30, 2020, WHO listed the novel coronavirus pneumonia outbreak as a Public...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12R1/93
CPCC12Q1/701C12Q1/686C12Q2600/16C12Q2600/166C12Q2561/101C12Q2537/143C12Q2545/101Y02A50/30
Inventor 郭圣明李丹金捷陆容迟大利吴大治夏懿
Owner SHANGHAI XINGYAO MED TECH DEV CO LTD
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