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

A Marburg virus, fluorescence quantitative technology, applied in the direction of fluorescence/phosphorescence, microbial measurement/inspection, biochemical equipment and methods, etc.

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

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Problems solved by technology

[0003] At present, the detection methods for Marburg virus include ELISA antigen detection, indirect immunofluorescence serological detection, virus isolation, RT-PCR nucleic acid detection, but the method of fluorescent quantitative PCR nucleic acid detection is recognized as a more intuitive and faster method. A detection method suitable for early diagnosis

Method used

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

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Embodiment Construction

[0032] The working principle of the new Marburg virus fluorescent quantitative PCR detection method of the present invention is to utilize the qualitative analysis of the change of the fluorescent signal to detect in real time the change of the amplification product amount of each cycle in the PCR amplification reaction, and to determine the difference by the relationship between the Ct value and the standard curve. The starting template was quantified. 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...

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Abstract

The invention discloses a novel Marburg virus fluorescent quantitative PCR (Polymerase Chain Reaction) detection method and a Marburg virus PCR detection system. The system consists of a primer, a probe, Premix Ex Tag reaction liquid and sterilized Tris water. The primer and the probe have high detection specificity and high sensitivity, are very suitable for the Marburg virus, and do not undergoany cross reaction with an Ebola virus.

Description

technical field [0001] The invention belongs to the field of biological detection, in particular to a rapid qualitative and quantitative detection method for Marburg virus and a PCR system for detection of Marburg virus, and 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 ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11G01N21/64
Inventor 杨宇王静韩辉胡孔新白琳徐宝梁
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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