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Novel fluorescence quantitative PCR (Polymerase Chain Reaction) detection method of encephalitis B virus and encephalitis B virus detection PCR system

A type of Japanese encephalitis virus, fluorescence quantitative technology, applied in fluorescence/phosphorescence, biochemical equipment and methods, determination/inspection of microorganisms, etc.

Inactive Publication Date: 2011-09-28
CHINESE ACAD OF INSPECTION & QUARANTINE
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Problems solved by technology

However, the method of fluorescent quantitative PCR nucleic acid detection has not been used yet, and it is a more intuitive, accurate and rapid detection standard suitable for early diagnosis

Method used

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  • Novel fluorescence quantitative PCR (Polymerase Chain Reaction) detection method of encephalitis B virus and encephalitis B virus detection PCR system
  • Novel fluorescence quantitative PCR (Polymerase Chain Reaction) detection method of encephalitis B virus and encephalitis B virus detection PCR system
  • Novel fluorescence quantitative PCR (Polymerase Chain Reaction) detection method of encephalitis B virus and encephalitis B virus detection PCR system

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Embodiment Construction

[0031] The working principle of the Japanese encephalitis virus fluorescent quantitative PCR detection method 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 amplification product amount of each cycle in the PCR amplification reaction, and to pass the Ct value and the standard curve. The relationship is quantified on the starting template. The present invention also relates to all the contents contained in the above-mentioned detection system.

[0032] The method includes the following steps:

[0033] 1) Design primer probes;

[0034] 2) Select fluorescein according to the instrument;

[0035] 3) Synthetic primers and probes;

[0036] 4) Prepare positive plasmid standard;

[0037] 5) run PCR;

[0038] 6) Data analysis;

[0039] 7) Extract viral RNA for system verification.

[0040] 1. Design of primers and probes

[0041] According to the name of the gene to be studied, Japanes...

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Abstract

The invention discloses a novel fluorescence quantitative PCR (Polymerase Chain Reaction) detection method of encephalitis B virus and an encephalitis B virus detection PCR system. The system consists of a primer and a probe, a Premix ex Taq reaction solution and sterilized Tris water. The primer and the probe have good detection specificity and high sensitivity; and the encephalitis B virus detection PCR system is very suitable for detecting encephalitis B virus and has no cross reaction on dengue fever virus and yellow fever 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 Japanese encephalitis virus and a PCR system for detection of Japanese encephalitis virus, and provides a pair of specific primers and probes. Background technique [0002] Epidemic encephalitis B (JE), referred to as JE, is an acute viral infectious disease mainly caused by central nervous system damage caused by the Japanese encephalitis virus, also known as Japanese encephalitis virus, transmitted by mosquitoes. It is widely distributed, including tropical, subtropical, temperate and mid-temperate regions, especially in some countries in Southeast Asia. The number of reported cases worldwide is more than 16,000 per year, with 5,000 deaths. At present, the diagnostic methods used for JE include ELISA serological detection, hemagglutination inhibition test (HI), RT-PCR nucleic acid detection and so on. However, the me...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11G01N21/64
CPCY02A50/30
Inventor 杨宇胡孔新王静姚李四白琳
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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