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Marburg and Ebola dual-virus fluorescent quantitative PCR (Polymerase Chain Reaction) detection method and system

A Marburg virus, Ebola virus technology, applied in fluorescence/phosphorescence, microbial assay/inspection, biochemical equipment and methods, etc.

Active Publication Date: 2011-08-03
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are ELISA antigen detection, indirect immunofluorescence serological detection, virus isolation, and RT-PCR nucleic acid detection methods for Marburg and Ebola virus detection methods, but the fluorescent quantitative PCR nucleic acid detection method is recognized as a more intuitive method. , a faster detection method suitable for early diagnosis

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  • Marburg and Ebola dual-virus fluorescent quantitative PCR (Polymerase Chain Reaction) detection method and system
  • Marburg and Ebola dual-virus fluorescent quantitative PCR (Polymerase Chain Reaction) detection method and system
  • Marburg and Ebola dual-virus fluorescent quantitative PCR (Polymerase Chain Reaction) detection method and system

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Embodiment Construction

[0042] Marburg of the present invention, the working principle of the new method of fluorescent quantitative PCR detection of Ebola double virus promptly utilizes the qualitative analysis of the change of fluorescent signal, detects in real time the change of the amplification product amount of each cycle in the PCR amplification reaction, through Ct value and The relationship to the standard curve was quantified against the starting template. The present invention also relates to all the contents contained in the above-mentioned detection system.

[0043] The method includes the following steps:

[0044] 1) Design primer probes;

[0045] 2) Select fluorescein according to the instrument;

[0046] 3) Synthetic primers and probes;

[0047] 4) Prepare positive plasmid standard;

[0048] 5) run PCR;

[0049] 6) Data analysis;

[0050] 7) Extract viral RNA for system verification.

[0051] 1. Design of primers and probes

[0052] According to the name of the gene to be stu...

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Abstract

The invention discloses Marburg and Ebola dual-virus fluorescent quantitative PCR (Polymerase Chain Reaction) detection method and system, wherein the detection system comprises primers, probes, a Premix EX Taq reaction solution and sterilizing Tris water. As two pairs of primers and probes have very good specificity, the detection system has high sensitivity and is suitable for simultaneously detecting Marburg and Ebola viruses without having cross reaction with other kinds of hemorrhagic fever arbovirus, such as yellow fever, dengue and rift valley fever.

Description

technical field [0001] The invention belongs to the field of biological detection, and in particular relates to a rapid qualitative and quantitative detection method for dual viruses of Marburg and Ebola, each of which provides a pair of specific primers and probes. Background technique [0002] Marburg virus (MbV), also known as green monkey disease virus and green monkey factor, is the first filovirus discovered by humans, which can cause high mortality in humans and other primates. Hemorrhagic fever, Marburg hemorrhagic fever has a sudden onset, often accompanied by severe headache and discomfort. Muscle pain is a common feature. The incubation period is generally 3 to 9 days, and the longer one can exceed 2 weeks. It can be used as a potential biological disease. The use of terrorist weapons or biological warfare agents is a virulent virus that requires the highest level of biological safety protection (P4 level). The WTO issued an alert saying, "Marburg virus is one of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68G01N21/64
CPCY02A50/30
Inventor 杨宇王静白琳胡孔新徐宝梁
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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