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Loop-mediated isothermal amplification detection primer set, reagent kit and method for SARS-CoV-2

A loop-mediated isothermal, detection kit technology, applied in the field of medical molecular biology diagnosis, can solve problems such as no SARS-COV-2 methods and kits, and achieve good specificity, protective activity, and guaranteed specificity. Effect

Pending Publication Date: 2020-06-23
JINAN CENTER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These advantages of LAMP have made it widely concerned since its invention, but the inventor found that in the medical field, only kits for detecting Mycobacterium tuberculosis, Salmonella and Legionella are commercialized, and there is no method and method for SARS-COV-2. Related reports about kits

Method used

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  • Loop-mediated isothermal amplification detection primer set, reagent kit and method for SARS-CoV-2
  • Loop-mediated isothermal amplification detection primer set, reagent kit and method for SARS-CoV-2
  • Loop-mediated isothermal amplification detection primer set, reagent kit and method for SARS-CoV-2

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Experimental program
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Effect test

Embodiment

[0087] 1. RNA extraction

[0088] The RNA used for amplification is extracted by a commercially available qualified extractor and reagents, without special requirements.

[0089] 2. Primer design and screening

[0090] According to the three gene sequences of RdRp, N, and E, multiple sets of primers were designed using Primer Explorer V5 software. Each primer set included a pair of specific inner primers and a pair of specific outer primers. The reaction process and results are monitored and screened to determine the best primers with fast response and high specificity. The screened primer sequences are listed in Table 1.

[0091] Table 1 Primer Sequence

[0092]

[0093]

[0094] 3. LAMP reaction system

[0095] The total volume of the LAMP reaction is 12.5 μL. During the reaction, 10 μL of the reaction solution is added, and then 2.5 μL of the RNA to be detected is added. A positive control and a negative control were also set up.

[0096] The positive control is...

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Abstract

The invention provides a loop-mediated isothermal amplification detection primer set, reagent kit and method for SARS-CoV-2, and belongs to the technical field of medical science and molecular biologydiagnosis. Based on stability and uniqueness of RdRp, E and N genes in the SARS-COV-2, three primer sets are designed to perform amplification detection on the gene, the primer concentration is reduced, the reaction specificity is improved, and the false positive is reduced. Therefore, the detection primer set, reagent kit and method, provided by the invention are high in detection speed, high insensitivity and high in specificity, and have favorable practical application value.

Description

technical field [0001] The invention belongs to the technical field of medical molecular biology diagnosis, and in particular relates to a SARS-CoV-2 ring-mediated isothermal amplification detection primer set, a kit and a detection method. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] Coronaviruses are a large family of viruses known to cause colds as well as more serious illnesses such as Middle East Respiratory Syndrome (MERS) and Severe Acute Respiratory Syndrome (SARS). SARS-COV-2 is a new strain of coronavirus that has never been found in humans before. Its gene sequence is 79.5% similar to SARS coronavirus, and the path of entry into cells is the same as that of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/701C12Q1/6844C12Q2531/119Y02A50/30
Inventor 汪运山杨炜华钊倩倩周云英徐祎慧王敏王军韩淑毅
Owner JINAN CENTER HOSPITAL
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