New fluorescence quantitative polymerase chain reaction (PCR) detection method for rift valley fever virus and rift valley fever virus detection PCR system

A Rift Valley Fever virus, fluorescence quantitative technology, applied in the direction of fluorescence/phosphorescence, determination/inspection of microorganisms, biochemical equipment and methods, etc.

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

[0003] At present, there are many detection methods for Rift Valley fever virus, such as virus isolation, agarose immunodiffusion, immunofluorescence staining, ELISA, virus neutralization test, hemagglutination inhibition test, enzyme-linked immunosorbent assay, etc., but Fluorescent quantitative PCR nucleic acid detection method is still recognized as a more intuitive and faster detection method suitable for early diagnosis

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  • New fluorescence quantitative polymerase chain reaction (PCR) detection method for rift valley fever virus and rift valley fever virus detection PCR system
  • New fluorescence quantitative polymerase chain reaction (PCR) detection method for rift valley fever virus and rift valley fever virus detection PCR system
  • New fluorescence quantitative polymerase chain reaction (PCR) detection method for rift valley fever virus and rift valley fever virus detection PCR system

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Embodiment Construction

[0032] The working principle of the new method for the detection of Rift Valley fever virus fluorescent quantitative PCR 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 through 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 studied, Rif...

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Abstract

The invention discloses a new fluorescence quantitative polymerase chain reaction (PCR) detection method for rift valley fever virus and a rift valley fever virus detection PCR system. The system consists of primers and a probe, Premix Ex Taq reaction solution, and sterile Tris water, wherein the primers and the probe have good specificity and high sensitivity, and are suitable for no cross reaction of the rift valley fever and Dengue 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 Rift Valley fever virus and a PCR system for detection of Rift Valley fever virus, and provides a pair of specific primers and probes. Background technique [0002] Rift Valley fever (Rift Valley fever, RVF), also known as sheep and cattle infectious enzootic hepatitis, is a deadly infectious disease prevalent in Africa. It infects humans and livestock through mosquito bites. The main symptoms are acute diarrhea and high fever, and then severe It can damage the liver and kidneys of humans and livestock, and some patients will die due to blood vessel rupture. The International Office of Epizootics (OIE) classifies it as a category A disease. The disease is more harmful to sheep, goats, and cattle, and can cause abortion in pregnant animals and a high proportion of newborn animals to die. Older non-pregnant animals are a...

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 杨宇王静胡孔新孙肖红白琳马雪征
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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