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Detection kit for ORF1a2 gene of Middle East respiratory syndrome coronavirus

A technology for respiratory syndrome and coronavirus, applied in the field of clinical medicine, can solve the problems of time-consuming, labor-intensive, cost, etc., and achieve the effect of simple method and broad application prospects.

Inactive Publication Date: 2015-11-11
黑龙江盈泰电气线路安全检测有限公司
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Problems solved by technology

[0006] The purpose of the present invention is to provide a MERS-CoV nucleic acid screening method based on loop-mediated isothermal amplification technology, in order to overcome the time-consuming, labor-intensive and high-cost defects of the prior art, and provide a simple, economical, fast and sensitive And specific MERS-CoV ORF1a2 gene detection kit

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  • Detection kit for ORF1a2 gene of Middle East respiratory syndrome coronavirus
  • Detection kit for ORF1a2 gene of Middle East respiratory syndrome coronavirus

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Embodiment Construction

[0035] The detection method of the differential diagnosis system shall be carried out in sequence as follows:

[0036] (1) Extraction of viral RNA: extract using traditional RNA extraction kits, or use extraction kits;

[0037] (2) Reverse transcription: according to the conventional reverse transcription method;

[0038](3) LAMP reaction: Prepare the above reaction system, mix the reaction system, and distribute it into 0.2ml PCR reaction tubes, 23 μl per tube, add the template for reverse transcription in step (2) into the PCR reaction tubes, each tube 2 μl, 2 μl of positive control substance and 2 μl of negative control substance, respectively, as positive control and negative control, mix well. Put it into a real-time turbidimeter or a constant temperature water bath, set the reaction conditions: 65°C, 60 minutes; 80°C, 5 minutes to stop the reaction, store at 4°C, add Smartgreen fluorescent dye to mix, develop under ultraviolet light (wavelength 302nm) ;

[0039] (4) J...

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Abstract

The invention relates to a detection kit for an ORF1a2 gene of Middle East respiratory syndrome coronavirus, and a loop-mediated isothermal amplification-based detection method for the Middle East respiratory syndrome coronavirus. The detection method is used for performing nucleic acid detection on the ORF1a2 gene of the Middle East respiratory syndrome coronavirus in a sample on the basis of the loop-mediated isothermal amplification principle; by using specific primers and utilizing a loop-mediated isothermal amplification platform to amplifying the target gene ORF1a2, under the assistance of a positive quality control and negative control and inner control detection system, screening or detection is performed on the Middle East respiratory syndrome coronavirus in the sample from a molecular level. The method disclosed by the invention has the characteristics of being simple, economic, rapid, sensitive and specific, is capable of overcoming the defects that the time and energy are wasted and the cost is high in the prior art, and has a wide application prospect.

Description

technical field [0001] The invention belongs to the field of clinical medicine, and relates to a biological diagnostic reagent, which relates to a Middle East respiratory syndrome coronavirus ORF1a2 (MERS-CoV) gene detection kit, which is a Middle East respiratory syndrome coronavirus nucleic acid based on a ring-mediated isothermal amplification technology screening method. Background technique [0002] Coronaviruses are a large class of viruses that can cause human diseases, and their clinical manifestations cover a wide range from the common cold to severe acute respiratory syndrome (SARS). Such viruses can also cause disease in many species of animals. At the end of 2012, a new coronavirus, never before seen in humans, was identified in residents of the Middle East. The virus, named Middle East Respiratory Syndrome Coronavirus (MERS-CoV), has now seen at least more than 50 laboratory-confirmed cases of human infection. At present, all cases of MERS-CoV infection have ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/6844C12Q1/701C12Q2531/119
Inventor 张文超孙嘉鹏赵赫
Owner 黑龙江盈泰电气线路安全检测有限公司
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