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Composition, kit and method for detecting SARS-CoV-2 and application of composition, kit and method

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

Active Publication Date: 2022-04-29
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 and method for detecting SARS-CoV-2 and application of composition, kit and method
  • Composition, kit and method for detecting SARS-CoV-2 and application of composition, kit and method
  • Composition, kit and method for detecting SARS-CoV-2 and application of composition, kit and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Embodiment 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] Embodiment 2, the 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 gel 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 RNA solution for direct detection.

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

[0088] System configuration:

[0089] According to the total reaction number N required for detection, add 18.5 μl RT-PCR amplification solution (MgCl 2 6mM, dNTP0.8mM, and 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 enzyme mixture (hot start Taq enzyme, reverse transcriptase, uracil ...

Embodiment 3

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

[0104] With primer and probe shown in embodiment 1, according to the method for embodiment 2, the pseudovirus of Omicron and Delta is detected, and experimental result is as follows Figure 1~3 shown. The results show that each channel can be detected normally, and the multiplex PCR system can detect the corresponding target and type Omicron and Delta.

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Abstract

The invention belongs to the field of molecular biological detection. In particular, the invention relates to the detection of SARS-CoV-2; more specifically, the invention relates to a composition, a kit and a method for detecting SARS-CoV-2 and application thereof. The invention provides a kit containing the composition, application of the composition and a method for detecting and typing the SARS-CoV-2 variant. By detecting variation sites with different characteristic functions, the Armike and Delta variants are subjected to typing, so that the typing detection of the SARS-CoV-2 variants is realized in a single-tube reaction system at the same time. Different strains can be treated differently, so that treatment and prevention are more efficient. The composition disclosed by the invention is low in cost, high in flux and simple and convenient to operate, and a result reading process can be judged through an amplification curve Ct value. The whole detection process is carried out under a single-tube closed condition, so that false positive and environmental pollution are avoided.

Description

technical field [0001] The invention belongs to the field of molecular biology detection; in particular, it relates to the detection of SARS-CoV-2; (Delta) detection. Background technique [0002] Novel coronavirus (2019-nCoV, SARS-CoV-2), on February 11, 2020, the International Virus Taxonomy Committee named the virus as SARS-CoV-2. Coronavirus is a large family of viruses. Before that, 6 species were known to infect humans, such as colds, Middle East Respiratory Syndrome (MERS) and Severe Acute Respiratory Syndrome (SARS). 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 family Coronaviridae (Coronavirus) in taxonomy, and has the characteristics of capsule and spike cytology The genome is a linear single-stranded positive-sense RNA ((+)ssRNA) virus. [0003] According to the official website of the World Health Organization, hundreds of variants of the new coronavirus have been di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2563/107C12Q2545/114Y02A50/30
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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