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Novel Ebola virus fluorescent quantitative PCR (Polymerase Chain Reaction) detection method and system

An Ebola virus, fluorescence quantitative technology, applied in the direction of fluorescence/phosphorescence, determination/inspection of microorganisms, biochemical equipment and methods, etc.

Inactive Publication Date: 2011-08-03
CHINESE ACAD OF INSPECTION & QUARANTINE
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Problems solved by technology

[0003] Currently, the detection methods for Ebola virus include immunohistochemical examination, serological examination, electron microscope examination, virus isolation, RT-PCR nucleic acid detection, and real-time RT-PCR nucleic acid detection. Each method has its own advantages and disadvantages. However, the fluorescent quantitative PCR nucleic acid detection method is recognized as a more intuitive and faster detection method suitable for early diagnosis

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  • Novel Ebola virus fluorescent quantitative PCR (Polymerase Chain Reaction) detection method and system
  • Novel Ebola virus fluorescent quantitative PCR (Polymerase Chain Reaction) detection method and system
  • Novel Ebola virus fluorescent quantitative PCR (Polymerase Chain Reaction) detection method and system

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Embodiment Construction

[0032] The working principle of the new method for Ebola virus fluorescent quantitative PCR detection of the present invention is to use the qualitative analysis of the change of the fluorescent signal to detect in real time the change of the amount of amplification product in each cycle in the PCR amplification reaction, and to pass the relationship between the Ct value and the standard curve Quantitative analysis of the starting template. The present invention also relates to all the contents contained in the above-mentioned detection system.

[0033] The method includes the following steps:

[0034] 1) Design primer probes;

[0035] 2) Select fluorescein according to the instrument;

[0036] 3) Synthetic primers and probes;

[0037] 4) Prepare positive plasmid standard;

[0038] 5) run PCR;

[0039] 6) Data analysis;

[0040] 7) Extract viral RNA for system verification.

[0041] 1. Design of primers and probes

[0042] According to the name of the gene to be studie...

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Abstract

The invention discloses Novel Ebola virus fluorescent quantitative PCR (Polymerase Chain Reaction) detection method and system, wherein the detection system comprises a primer, a probe, a Premix EX Taq reaction solution and sterilizing Tris water. As the primer and the probe has good detection specificity, the detection system has high sensitivity and is suitable for detecting Ebola viruses without having cross reaction with Marburg viruses.

Description

technical field [0001] The invention belongs to the field of biological detection, in particular to a rapid qualitative and quantitative detection method for Ebola virus, and provides a pair of specific primers and probes. Background technique [0002] Ebola virus (EBOV) can cause a zoonotic infectious disease, namely Ebola hemorrhagil fever (EBHF), in 1976 in southern Sudan and Zaire, which is now Congo (Kinshasa) The first outbreak occurred in the Ebola River area in the United States, and the mortality rate of patients was as high as 90%. The Ebola virus is highly contagious and can be transmitted through direct contact with the infected person's body fluids or other infected tissues, skin ulcers and droplets. Infected individuals experience severe bleeding and can develop shock syndrome. The World Health Organization has listed EBOV as one of the most harmful viruses to humans, that is, level 4 viruses, and related experimental operations must be carried out in P4 leve...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11G01N21/64
Inventor 杨宇王静白琳胡孔新孙肖红徐宝梁
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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