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Novel freeze-drying detection reagent for coronavirus nucleic acid and preparation method thereof

A coronavirus and detection reagent technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, DNA/RNA fragments, etc., can solve the problems affecting the stability and inactivation of active components, so as to achieve convenient operation and reduce The effect of error and low cost of raw materials

Pending Publication Date: 2022-07-15
SUZHOU GENO TRUTH BIOTECHNOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, during the freeze-drying and storage process of the novel coronavirus nucleic acid detection reagent, many factors (chemical composition, freezing rate, freezing and dehydration stress, glass transition temperature, residual moisture in the dry solid, temperature and humidity of the storage environment, etc.) Affect the stability of the active components and even lead to inactivation

Method used

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  • Novel freeze-drying detection reagent for coronavirus nucleic acid and preparation method thereof
  • Novel freeze-drying detection reagent for coronavirus nucleic acid and preparation method thereof
  • Novel freeze-drying detection reagent for coronavirus nucleic acid and preparation method thereof

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preparation example Construction

[0038] One aspect of the embodiments of the present invention provides a method for preparing a freeze-dried detection reagent for novel coronavirus nucleic acid, comprising: adding hot-start Taq DNA polymerase (HS Taq) and / or hot-start Fast Taq DNA polymerase ( HSFast Taq), reverse transcription polymerase (M-MLV), Taq PCR Buffer (Taq PCR Buffer) and / or Fast Taq PCR Buffer (Fast Taq PCR Buffer), deoxyribonucleotide triphosphate (dNTP), frozen The dry protective agent and the primer set are thoroughly mixed to form a mixture, which is then freeze-dried to prepare a freeze-dried detection reagent for novel coronavirus nucleic acid.

[0039] The hot-start Taq DNA polymerase in the present invention is an enzyme that is independently induced, expressed, separated, purified and modified by Suzhou Industrial Park Longzhao Biotechnology Research Institute Co., Ltd., and can be used in a freeze-drying system. Commercial enzymes on the market contain certain These substances should no...

Embodiment 1

[0104] Example 1: Single and novel coronavirus nucleic acid detection reagent

[0105] 1. The components of the freeze-dried protective agent are shown in Table 1:

[0106] Table 1 lyophilized protective agent formula: 2.5 × lyophilized Mix1

[0107]

[0108] 2. Preparation of freeze-dried detection reagents for novel coronavirus nucleic acid:

[0109] Table 2 Components of lyophilized detection reagents

[0110] The components of freeze-dried detection reagents are shown in Table 2:

[0111]

[0112] The components of the freeze-dried detection reagent were mixed, and then freeze-dried to obtain a freeze-dried detection reagent for novel coronavirus nucleic acid. segments: 10;

[0113] Table 3 Freeze drying process

[0114]

[0115] 3. Add 20 μL of the above-prepared freeze-dried detection reagent, the sample to be tested and water, mix well, and then perform the HS-Taq One ​​Step RT-PCR reaction. The specific steps are shown in Table 4:

[0116] Table 4 HS-Taq...

Embodiment 2

[0120] Example 2: Single novel coronavirus nucleic acid detection reagent

[0121] 1. The components of the freeze-dried protective agent are shown in Table 5:

[0122] Table 5 lyophilized protective agent formula: 2.5 × lyophilized Mix2

[0123]

[0124] 2. Preparation of freeze-dried detection reagents for novel coronavirus nucleic acid:

[0125] Table 6 Components of lyophilized detection reagents

[0126] The components of freeze-dried detection reagents are shown in Table 6:

[0127]

[0128] The components of the freeze-dried detection reagent are mixed, and then freeze-dried to obtain a freeze-dried detection reagent for novel coronavirus nucleic acid. segment: 11;

[0129] Table 7 Freeze drying process

[0130]

[0131] 3. Add 20 μL of the lyophilized detection reagent prepared above, the sample to be tested and water, mix well, and then perform the HS-Taq One ​​Step RT-PCR reaction. The specific steps are shown in Table 8:

[0132] Table 8 HS-Taq One ​​...

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Abstract

The invention discloses a novel freeze-dried detection reagent for coronavirus nucleic acid and a preparation method of the novel freeze-dried detection reagent. The preparation method comprises the following steps: fully mixing a hot start Taq DNA polymerase and / or a hot start Fast Taq DNA polymerase, a reverse transcription polymerase, a Taq PCR buffer solution and / or a Fast Taq PCR buffer solution, deoxyribonucleotide triphosphate, a freeze-drying protective additive and a primer group to form a mixture, and then freeze-drying to obtain the novel freeze-drying detection reagent for coronavirus nucleic acid. When the novel freeze-dried detection reagent for coronavirus nucleic acid prepared by the invention is used, only an actual sample or a template and water need to be added, so that the workload of an operator and an operation error are greatly reduced; meanwhile, the detection reagent has the advantages of high stability, easiness in transportation, storage resistance, convenience in operation, more ideal experimental results and the like, and the storage life can be up to 3 years.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid detection, and in particular relates to a freeze-dried detection reagent for novel coronavirus nucleic acid and a preparation method thereof. Background technique [0002] Novel coronavirus pneumonia is a disease caused by the novel coronavirus (SARS-CoV-2). At present, the source of infection is mainly patients infected with the new coronavirus, and the latent infection may also become the source of infection. It is mainly transmitted through respiratory droplets and contact transmission, and the population is generally susceptible. The novel coronavirus belongs to the genus betacoronavirus. [0003] Polymerase Chain Reaction (PCR) is a molecular biology technique used to amplify specific DNA fragments. When DNA is denatured at high temperature in vitro, it will become single-stranded. At low temperature, the primer and single-strand are based on the principle of complementary base pairing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2563/107C12Q2545/114C12Q2521/107
Inventor 张惠丹吴宁
Owner SUZHOU GENO TRUTH BIOTECHNOLOGY CO LTD
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