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A new method for the detection of yellow fever virus by fluorescent quantitative PCR and a PCR system for the detection of yellow fever virus

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

Active Publication Date: 2011-12-14
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Abstract
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Problems solved by technology

At present, the detection methods used for yellow fever virus include microwell hybridization, chip detection, and nested PCR. , a detection method suitable for early diagnosis

Method used

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  • A new method for the detection of yellow fever virus by fluorescent quantitative PCR and a PCR system for the detection of yellow fever virus
  • A new method for the detection of yellow fever virus by fluorescent quantitative PCR and a PCR system for the detection of yellow fever virus
  • A new method for the detection of yellow fever virus by fluorescent quantitative PCR and a PCR system for the detection of yellow fever virus

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Embodiment Construction

[0033] The working principle of the new method for fluorescent quantitative PCR detection of yellow fever virus of the present invention is: utilize the change of fluorescence signal, qualitative analysis, real-time detection of the change of each cycle amplification product amount in the PCR amplification reaction, through the relation of Ct value and standard curve Quantitative analysis of the starting template.

[0034] The inventive method comprises the following steps:

[0035] 1) Design primer probes;

[0036] 2) Select fluorescein according to the instrument;

[0037] 3) Synthetic primers and probes;

[0038] 4) Prepare positive plasmid standard;

[0039] 5) run PCR;

[0040] 6) Data analysis;

[0041] 7) Extract viral RNA for system verification.

[0042] 1. Design of primers and probes

[0043] According to the name of the gene to be studied, Yellow Fever Virus (YFV), the corresponding whole gene sequence was found in genebank (www.ncbi.nlm.nih.gov), and DNASTA...

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Abstract

The invention discloses a novel fluorescence quantitative PCR (Polymerase Chain Reaction) detection method for yellow fever viruses and a yellow fever virus detection PCR system. The yellow fever virus detection PCR system comprises primers and probes, a Premix Ex Taq reaction liquid, and sterilized Tris water, wherein the primers and the probes are good in detection specificity and high in sensitivity, is very suitable to bouquet fever viruses, and has no cross reactions with the yellow fever viruses and encephalitis B 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 yellow fever virus and a PCR system for detection of yellow fever virus, and provides a pair of specific primers and probes. Background technique [0002] Yellow fever is an acute viral disease and an important cause of hemorrhagic disease. Every year there are at least 200,000 cases of yellow fever worldwide and 30,000 deaths. Due to the high mortality rate and strong infectivity of yellow fever, it has been included as one of the quarantine infectious diseases stipulated by the World Health Organization. Some national laws require travelers to endemic areas to be vaccinated. Yellow fever is one of only a few diseases that has this "honor". Yellow fever virus belongs to group B of arboviruses and is an RNA virus with a spherical shape and a size of about 20-60 nanometers. At present, the detection methods used for ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11G01N21/64
Inventor 杨宇王静胡孔新白琳张晓龙
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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