Fully-premixed freeze-drying multi-fluorescent PCR detection kit for novel coronavirus, influenza A virus and influenza B virus and detection method thereof

A type of influenza B virus and detection kit technology, which is applied in the field of in vitro diagnosis and biological detection, can solve the problems of time consumption and impact on virus detection rate, increase stability, avoid repeated freezing and thawing and multiple packaging, The effect of a low level of operation

Pending Publication Date: 2021-05-07
青岛巴特菲科技发展有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the currently commonly used method is to first detect the new coronary pneumonia after sampling the patients, and then detect ea...

Method used

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  • Fully-premixed freeze-drying multi-fluorescent PCR detection kit for novel coronavirus, influenza A virus and influenza B virus and detection method thereof
  • Fully-premixed freeze-drying multi-fluorescent PCR detection kit for novel coronavirus, influenza A virus and influenza B virus and detection method thereof
  • Fully-premixed freeze-drying multi-fluorescent PCR detection kit for novel coronavirus, influenza A virus and influenza B virus and detection method thereof

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Experimental program
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Effect test

Embodiment 1

[0039] 1. The lyophilized solid-state RT-PCR Mix contains primer sets and probes corresponding to the N gene of SARS-CoV-2, the M gene of influenza A, the M gene of influenza B, and the RNAse P primer sequence of the internal reference gene of human origin. The solid-state RT-PCR Mix mainly contains the N gene of the new coronavirus (SARS-CoV-2), the M gene of the influenza A virus, the M gene of the influenza B virus and the specificity of the genome of the internal standard gene RNaseP before lyophilization. Primers and probes, dNTP, Taq DNA polymerase, reverse transcriptase, RNase inhibitor, UDG enzyme and freeze-dried compound protective agent, and the balance is DEPC treated water. The specific components and contents are shown in Table 2.

[0040] Table 2: Novel Coronavirus and Influenza A, Influenza B Virus RT-PCRMix Liquid

[0041]

[0042]

[0043] Preparation steps of freeze-dried solid-state novel coronavirus and influenza A and influenza B virus RT-PCR mix pa...

Embodiment 2

[0072] The operation method of embodiment 2 detection kit

[0073] The operation method of the freeze-dried rapid fluorescent quantitative PCR detection kit for human new coronavirus, type A and type B influenza virus:

[0074] Including the following steps:

[0075] (1) Template RNA extraction, using a nucleic acid extraction kit to extract RNA from the sample to be tested.

[0076] (2) Prepare reagents: add 15 μL of reconstituted Buffer RB to each PCR tube with freeze-dried particles, and add 50 μl of reconstituted Buffer RB to the freeze-dried vials of positive control and negative control;

[0077] (3) Adding samples and controls: According to the number of experimental samples, add 5ul of the extracted sample RNA to each of the above PCR tubes, and take 5ul of the negative and positive controls after reconstitution, and add them to the corresponding PCR tubes. Put the cap on the PCR tube.

[0078] (4) Centrifuge and put it into a fluorescent quantitative PCR instrument...

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Abstract

The invention discloses a multi-fluorescent PCR rapid detection kit for novel coronavirus, influenza A virus and influenza B virus. The kit comprises freeze-dried solid RT-PCR Mix, liquid redissolution Buffer, freeze-dried solid positive control and freeze-dried solid negative control, wherein the freeze-dried solid RT-PCR Mix contains a primer group corresponding to primer sequences of a SARS-CoV-2 specific gene ORF1ab, an influenza A M gene, an influenza B M gene and a human reference gene RNAse P. The kit combines a multi-fluorescent quantitative PCR technology and a freeze-drying process, utilizes three pairs of special primers and human reference genes to amplify specific sequences of three pathogens in vitro, and performs real-time detection in combination with a fluorescent probe. The detection method is simple and convenient to operate, has low requirements for the operation level of detection personnel, and can detect three common respiratory pathogens at a time, the detection time and the detection cost are greatly saved, rapid screening of large-batch samples is realized, the whole detection process only takes 40 minutes to 1 hours, and results are accurate and reliable.

Description

technical field [0001] The invention belongs to the technical field of in vitro diagnosis and biological detection, and in particular relates to a fully premixed freeze-dried multiple fluorescent PCR detection kit for novel coronavirus, type A and type B influenza viruses and a detection method thereof. Background technique [0002] Coronavirus disease 2019 (Corona Virus Disease 2019, COVID-19) is an acute respiratory infectious disease caused by a new coronavirus (new Corona Virus, SARS-CoV-2). SARS-CoV-2 belongs to the new type of coronavirus belonging to the genus β. It is an RNA virus with an envelope. The particles are round or oval, often pleomorphic, with a diameter of 60-140nm. It is currently known to infect humans. The seventh coronavirus, the remaining six coronaviruses are HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV and MERS-CoV. Among them, the first four are more common in the population, with low pathogenicity, and generally only cause mild respirato...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/701C12Q2600/166C12Q2563/107C12Q2545/114C12Q2537/143Y02A50/30
Inventor 李然栋马清霞薄慧文
Owner 青岛巴特菲科技发展有限公司
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