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Novel coronavirus nucleic acid PCR-colloidal gold immunochromatography detection kit

A technology of immunochromatography and detection kit, which is applied in the field of novel coronavirus nucleic acid colloidal gold immunochromatography detection kit, can solve problems such as inability to effectively monitor the nucleic acid detection process, inability to set internal reference gene detection, and contamination. Achieve the effects of saving detection time, high clinical applicability, and high precision

Inactive Publication Date: 2021-07-30
北京华诺奥美医学检验实验室有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] During the whole experiment process, sample nucleic acid extraction and PCR amplification are required, and contamination or other interference factors may occur. Traditional colloidal gold only has detection lines and quality control lines, and cannot set internal reference gene detection, and cannot effectively monitor the entire nucleic acid detection process. , may cause false negative results and interfere with judgment

Method used

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  • Novel coronavirus nucleic acid PCR-colloidal gold immunochromatography detection kit
  • Novel coronavirus nucleic acid PCR-colloidal gold immunochromatography detection kit
  • Novel coronavirus nucleic acid PCR-colloidal gold immunochromatography detection kit

Examples

Experimental program
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Effect test

Embodiment 1

[0036] A novel coronavirus nucleic acid PCR-colloidal gold immunochromatography detection kit, comprising: amplification reaction solution (i.e. amplification reaction MIX, purchased from Nanjing Nuoweizan Biotechnology Co., Ltd. (article number: Q222-CN), the The reaction system includes reverse transcriptase, Taq DNA polymerase, 4 kinds of dNTP and various ions required for PCR amplification reaction solution), CoV detection solution, positive control (mixture of virus target gene plasmid and internal reference plasmid), negative control product (RNase Free ddH2O) and chromatographic strip (coated with colloidal gold particles and antibodies); the CoV detection solution includes specific marker primers for the new coronavirus open reading frame 1ab and the gene encoding the nucleocapsid protein N, internal reference genes RNase P-specific labeled primers and RNase Free ddH 2 O.

[0037] The primer sequences in this example are as follows:

[0038] New coronavirus amplifica...

Embodiment 2

[0073] Performance verification of the kit described in embodiment 1

[0074] (1) Sample processing

[0075] Select the national reference product of the new coronavirus nucleic acid detection reagent purchased to verify the performance of the kit, take the sample to be tested into a 1.5mL centrifuge tube, use the nucleic acid extraction or purification reagent produced by Tiangen Biochemical Technology (Beijing) Co., Ltd., and strictly follow the instructions. RNA extraction operation. Extracted RNA samples should be tested immediately or stored below -70°C and avoid repeated freezing and thawing.

[0076] (2) PCR amplification

[0077] Configure the PCR amplification reaction solution (20 μL for each reaction) according to Table 3.

[0078] Table 3 PCR amplification reaction solution

[0079] components 1 reaction volume CoV detection solution 7.5μL Amplification reaction solution 12.5μL total capacity 20 μL

[0080] Aliquot the prepa...

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Abstract

The invention discloses a novel coronavirus nucleic acid PCR-colloidal gold immunochromatography detection kit. The kit comprises an amplification reaction solution, a CoV detection solution, a positive reference substance and a negative reference substance, and the CoV detection liquid comprises a specific marker primer aiming at a new coronavirus open reading frame 1ab and a coding nucleocapsid protein N gene, a specific marker primer of a reference gene RNase P and RNase Free ddH2O. According to the kit, after nucleic acid is extracted from a collected cell sample, a specific fragment is amplified under the action of reverse transcriptase and Taq DNA polymerase, then the nucleic acid detection of the new coronavirus is realized by utilizing colloidal gold immunochromatography, in addition, the kit adopts a reference gene RNase P to carry out tracking quality control on sample collection, nucleic acid extraction and PCR amplification processes, and the whole experiment process is ensured to be within a controllable range.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a novel coronavirus (COVID-19) nucleic acid colloidal gold immunochromatography detection kit. Background technique [0002] 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). The novel coronavirus is a new strain of coronavirus that has never been found in humans before. [0003] The main transmission routes of the new coronavirus are respiratory droplet transmission and contact transmission, while aerosol and feces-oral transmission routes need to be further clarified. Epidemiological investigations show that most cases can be traced to close contacts with confirmed cases. [0004] Common signs after a person is infected with a coronavirus include respiratory symptoms, fever, cough, shortness of breath, and dyspnea. In mor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6804C12R1/93
CPCC12Q1/701C12Q1/6804C12Q2600/166
Inventor 于超计吴文立王倩玉魏星
Owner 北京华诺奥美医学检验实验室有限公司
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