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Visual LAMP synchronous detection kit and detection method for SARS-CoV-2

A virus detection and ring-mediated isothermal technology, applied in the field of visual LAMP simultaneous detection kits, can solve the problems of long detection time, high cost and high test cost

Pending Publication Date: 2021-10-12
INST OF MICROBIOLOGY JIANGXI ACADEMY OF SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, more expensive instruments and professional technicians are required, the detection time is long, and the test cost is also high

Method used

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  • Visual LAMP synchronous detection kit and detection method for SARS-CoV-2
  • Visual LAMP synchronous detection kit and detection method for SARS-CoV-2

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Embodiment 1

[0017] In the present invention, the method for using the kit preferably includes the following steps: 1 μL of genomic DNA to be detected, 0.5 μL of SARS-CoV-2-N-F3 primer at a concentration of 100 μM, and SARS-CoV-2 at a concentration of 100 μM - 0.5 μL of N-B3 primer, 2 μL of SARS-CoV-2-N-FIP primer at a concentration of 100 μM, 2 μL of a SARS-CoV-2-N-BIP primer at a concentration of 100 μM, 1×Lamp PCR Master Mix 12.5 μL, 0.16 U / μL of Bst 3.0 DNA polymerase 0.5 μL, ddH 2 O to make up to 25 μL.

[0018] Amplification results such as figure 1 As shown, 1 in the figure is the LAMP synchronous detection result of SARS-CoV-2 in this embodiment, M is DNA Marker, 1: SARS-CoV-2 gene reverse transcription plasmid; 2: double distilled water (negative control) by As can be seen from the figure, typical trapezoidal bands appeared in the positive group.

Embodiment 2

[0020] In the present invention, the method for using the kit preferably includes the following steps: 1 μL of genomic DNA to be detected, 0.5 μL of SARS-CoV-2-N-F3 primer at a concentration of 100 μM, and SARS-CoV-2 at a concentration of 100 μM - 0.5 μL of N-B3 primer, 2 μL of SARS-CoV-2-N-FIP primer at a concentration of 100 μM, 2 μL of a SARS-CoV-2-N-BIP primer at a concentration of 100 μM, 1×Lamp PCR Master Mix 12.5 μL, 0.16 U / μL of Bst 3.0 DNA polymerase 0.5 μL, Calcein at a concentration of 625 μM, Mn at a concentration of 12.5 mM 2+ , ddH 2 O to make up to 25 μL.

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Abstract

The invention discloses a visual reverse transcription loop-mediated isothermal amplification kit for detecting a novel coronavirus and a use method thereof. The kit is composed of a novel coronavirus nucleocapsid (N) gene loop-mediated isothermal amplification primer group, and the kit is mainly composed of a novel coronavirus N gene loop-mediated isothermal amplification primer group solution, a loop-mediated isothermal amplification reaction mixed solution, a calcein-Mn2+ dye mixed solution, Bst 3.0DNA polymerase and a novel coronavirus N gene positive quality control product. The novel coronavirus can be quickly detected by adopting a visual reverse transcription loop-mediated isothermal amplification detection method. The method can complete detection within 70 minutes, has strong specificity and high sensitivity, and provides a simple and fast way for prevention and control of the novel coronavirus and investigation and analysis of the epidemic trend.

Description

technical field [0001] The invention belongs to the technical field of molecular detection, and in particular relates to a visual LAMP synchronous detection kit and detection method for SARS-CoV-2. Background technique [0002] The new coronavirus, also known as the new coronavirus, has been named "SARS-CoV-2" by the International Committee on Taxonomy of Viruses. The clinical manifestations of novel coronavirus pneumonia are mainly fever, fatigue, and dry cough, and upper respiratory symptoms such as nasal congestion and runny nose are rare. About half of the patients developed dyspnea more than a week later, and severe cases rapidly progressed to acute respiratory distress syndrome, septic shock, metabolic acidosis that was difficult to correct, and coagulation dysfunction. [0003] According to the latest guidelines, the use of fluorescent RT-PCR to detect positive nucleic acid of the new coronavirus is still the gold standard for diagnosis. However, more expensive inst...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6844C12Q2521/101C12Q2531/119
Inventor 靳亮马广强关丽梅黄旭春占智高况文东王金昌陈俊晖周溪徐新平
Owner INST OF MICROBIOLOGY JIANGXI ACADEMY OF SCI
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