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Detection kit for simultaneously detecting nine respiratory tract viruses including SARS-CoV-2

A detection kit, sars-cov-2 technology, applied in the direction of microorganism-based methods, microorganism measurement/testing, microorganisms, etc., can solve the problems of large demand and achieve high sensitivity, good specificity, and low cost Effect

Inactive Publication Date: 2020-08-28
SHANDONG ACV BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is a large demand for novel coronavirus-related detection products, and rapid identification of novel coronavirus infection or other common respiratory virus infections has become an urgent problem to be solved by clinical and public health departments

Method used

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  • Detection kit for simultaneously detecting nine respiratory tract viruses including SARS-CoV-2
  • Detection kit for simultaneously detecting nine respiratory tract viruses including SARS-CoV-2
  • Detection kit for simultaneously detecting nine respiratory tract viruses including SARS-CoV-2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 is used to specifically detect the primer set and fluorescent probe of novel coronavirus SARS-CoV-2

[0062] Download the sequence of the new coronavirus SARS-CoV-2 published in the GISAID database, use the MAFFT analysis software to compare the sequences, and analyze the conserved regions of its genome. Dozens of specific primers and probe sequences for the detection of the novel coronavirus ORF1ab gene were designed in these conserved regions.

[0063] The design of primers and probes follows the following principles:

[0064] Choose the probe first, and then design the primers to be as close to the probe as possible. The selected sequence should be highly specific, and try to choose an amplified fragment with minimal secondary structure; this is because secondary structure will affect the efficiency of the reaction, and it will also hinder the amplification of the enzyme. The secondary structure detection is carried out first. If the secondary structure c...

Embodiment 2

[0090] Embodiment 2 simultaneously detects the detection kit of nine kinds of respiratory viruses that comprise SARS-CoV-2

[0091] A detection kit for simultaneous detection of nine respiratory viruses including SARS-CoV-2, including detection of influenza A virus, influenza B virus, new coronavirus SARS-CoV-2, rhinovirus, parainfluenza virus, respiratory syndrome Primer sets and probe sets for cellular virus, coronavirus 229E subtype, coronavirus OC43 subtype, coronavirus HKU1 subtype; and reaction solutions, enzyme mixtures, positive quality controls, and negative quality controls.

[0092] specifically,

[0093] The sequence of the upstream primer for detecting influenza A virus is shown in SEQ ID No.1, the sequence of the downstream primer is shown in SEQ ID No.2, and the sequence of the fluorescent probe is shown in SEQ ID No.3;

[0094] The sequence of the upstream primer for detecting influenza B virus is shown in SEQ ID No.4, the sequence of the downstream primer is ...

Embodiment 3

[0123] Embodiment 3 adopts the method for the kit of embodiment 2 to detect nine kinds of respiratory viruses simultaneously

[0124] 1. To extract the nucleic acid of the sample to be tested, use the nucleic acid extraction kit (magnetic bead method) of Shandong Aikewei Biotechnology Co., Ltd. (Luji Machinery No. 20190256).

[0125] 2. Prepare the reaction system, prepare the reaction solution, primer-probe mixture, and enzyme mixture in the kit according to the kit instructions.

[0126] Table 3 shows the preparation methods for detecting influenza A virus, influenza B virus, and novel coronavirus system.

[0127] Table 3 Detection of influenza A virus, influenza B virus, novel coronavirus SARS-CoV-2 system preparation method

[0128] Reaction system components Additional amount (μL) / per person Enzyme Mix 4 The reaction solution 12 Primer Probe Mix A 4 total capacity 20

[0129] The preparation method of the detection system for rhin...

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Abstract

The invention discloses a detection kit for simultaneously detecting nine respiratory tract viruses including SARS-CoV-2, and belongs to the technical field of biological detection. The kit comprisesprimer groups and fluorescent probes of flu A viruses, flu B viruses, novel coronaviruses, nasal viruses, para-flu viruses, respiratory tract syncytial viruses, coronavirus 229E subtype, coronavirus OC43 subtype and coronavirus HKU1 subtype, as well as a reaction solution, an enzyme mixed solution, a positive quality control product and a negative quality control product. Rapid confirmation and screening of the nine common respiratory tract viruses including the novel coronavirus SARS-CoV-2 can be achieved, meanwhile, a real-time fluorescent quantitative PCR technology is used, and the kit ishigh in sensitivity and good in specificity.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a detection kit for simultaneously detecting nine respiratory viruses including SARS-CoV-2. Background technique [0002] Polymerase Chain Reaction (Polymerase Chain Reaction) is catalyzed by DNA polymerase, using the parent strand DNA as a template, and using a specific primer as the starting point of extension, through steps such as denaturation, annealing, and extension, in vitro replicates DNA that is complementary to the parent strand template DNA The process of the sub-strand DNA. DNA polymerase uses single-stranded DNA as a template to initiate synthesis with a small piece of double-stranded DNA. One or two artificially synthesized oligonucleotide primers combine with a complementary sequence in the single-stranded DNA template to form a partial double strand. Under suitable temperature and environment, DNA polymerase will add deoxygenated mononucleotide to t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12R1/93
CPCC12Q1/701C12Q1/6851C12Q2600/16C12Q2600/158C12Q2563/107C12Q2561/113C12Q2531/113C12Q2537/143
Inventor 王海岩张通邓雪萍刘奕李艳艳陈旭赵书清张国宁
Owner SHANDONG ACV BIOTECH CO LTD
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