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Composition, kit, method for detecting sars-cov-2 and use thereof

A sars-cov-2, composition technology, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve problems such as non-existing detection

Active Publication Date: 2022-07-05
深圳联合医学检验实验室
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, there is no reagent for detecting Omicron and delta mutant strains and for distinguishing BA.1 and BA.2

Method used

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  • Composition, kit, method for detecting sars-cov-2 and use thereof
  • Composition, kit, method for detecting sars-cov-2 and use thereof
  • Composition, kit, method for detecting sars-cov-2 and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Example 1. Primers and probes used in the present invention

[0080] Table 1

[0081]

[0082] Wherein, the fluorescent reporter group of SEQ ID NO:3 is FAM; the fluorescent reporter group of SEQ ID NO:6 is VIC; the fluorescent reporter group of SEQ ID NO:9 is ROX; the fluorescent reporter group of SEQ ID NO:12 The group is CY5.

Embodiment 2

[0083] Example 2. Method for detecting SARS-CoV-2

[0084] Specimen type: oropharyngeal swab, nasopharyngeal swab.

[0085] Nucleic acid extraction:

[0086] Use commercial RNA extraction kits, such as nucleic acid extraction reagents based on silica membrane spin column method or nucleic acid extraction reagents based on magnetic bead method, and operate according to the kit instructions, and finally collect 80 μL of RNA solution for direct detection.

[0087] Or store at -80°C. Both negative and positive controls need to be extracted.

[0088] System configuration:

[0089] According to the total number of reactions N required for detection, add 18.5 μl RT-PCR amplification solution (MgCl) to each tube of PCR 2 6mM, dNTP 0.8mM, upstream primer / downstream primer concentration is 0.2μM, probe concentration is 0.25μM, universal upstream primer 0.6μM, universal downstream primer 0.6μM) and 1.5 μl of enzyme mixture (hot start Taq enzyme, reverse transcriptase, uracil glycos...

Embodiment 3

[0103] Embodiment 3, the detection result of the composition test sample of the present invention

[0104] The primers and probes shown in Example 1 were used to detect the pseudoviruses of Omicron and Delta according to the method of Example 2, and the experimental results were as follows: Figures 1~3 shown. The results showed that each channel could be detected normally, and the multiplex PCR system could detect the corresponding targets and type Omicron and Delta.

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Abstract

The invention belongs to the field of molecular biology detection; in particular, it relates to the detection of SARS-CoV-2; more specifically, it relates to a composition, kit, method and use for the detection of SARS-CoV-2. The present invention provides a kit comprising the composition, use of the composition, and a method for detecting and typing SARS-CoV-2 variant strains. By detecting different characteristic functional variation sites, Omicron and Delta mutants were typed, so as to simultaneously realize the detection of SARS-CoV-2 mutant typing in a single-tube reaction system. This enables different strains to be treated differently, making treatment and prevention more efficient. The composition of the invention has low cost, high throughput and simple operation, and the result reading process can be determined by the Ct value of the amplification curve. The whole process of detection is carried out under single-tube closed conditions to avoid false positives and environmental pollution.

Description

technical field [0001] The invention belongs to the field of molecular biology detection; specifically, it relates to the detection of SARS-CoV-2; more specifically, it relates to SARS-CoV-2 variant strains Omicron BA.1, BA.2 and Delta (Delta) detection. Background technique [0002] The new coronavirus (2019-nCoV, SARS-CoV-2) was named SARS-CoV-2 by the International Committee on Taxonomy of Viruses on February 11, 2020. Coronavirus is a large virus family, 6 species are known to infect humans before, such as can cause colds and Middle East respiratory syndrome (MERS) and severe acute respiratory syndrome (SARS), etc., while SARS-CoV-2 It is a new strain of coronavirus that has never been found in humans before. The virus belongs to the β-type coronavirus in the genus Coronaviridae (Coronaviridae) in taxonomy, with envelope and spike cytology characteristics The genome is a linear single-stranded positive-stranded RNA ((+)ssRNA) virus. [0003] According to the official ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2563/107C12Q2545/114Y02A50/30
Inventor 谭爱女郭永超王艳平蔡锦刚徐露
Owner 深圳联合医学检验实验室
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