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New chikungunya virus fluorescence quantitative polymerase chain reaction detection method and chikungunya virus polymerase chain reaction detection system

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

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

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Problems solved by technology

At present, the detection methods for Chikungunya virus include ELISA, immunochromatography, immunofluorescence antibody assay (IFA), virus isolation, RT-PCR and Real-time PCR and other nucleic acid amplification methods, and fluorescent quantitative PCR nucleic acid detection methods. The method is recognized as a more intuitive and faster detection standard suitable for early diagnosis

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  • New chikungunya virus fluorescence quantitative polymerase chain reaction detection method and chikungunya virus polymerase chain reaction detection system
  • New chikungunya virus fluorescence quantitative polymerase chain reaction detection method and chikungunya virus polymerase chain reaction detection system
  • New chikungunya virus fluorescence quantitative polymerase chain reaction detection method and chikungunya virus polymerase chain reaction detection system

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Embodiment Construction

[0031] The working principle of the new method for the detection of Chikungunya virus by fluorescence quantitative PCR is to use the qualitative analysis of changes in fluorescence signals to detect the changes in the amount of amplified products in each cycle in the PCR amplification reaction in real time. The relationship is quantitatively analyzed on the starting template. The 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) Choose fluorescein according to the instrument;

[0035] 3) Synthesis of primers and probes;

[0036] 4) Prepare positive plasmid standards;

[0037] 5) Run PCR;

[0038] 6) Data analysis;

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

[0040] 1. Design of primer probe

[0041] According to the name of the gene to be studied, Chikungunya virus (Chikungunya virus), find the corresponding full gene sequence in genebank (...

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Abstract

The invention discloses a new chikungunya virus fluorescence quantitative polymerase chain reaction (PCR) detection method and a chikungunya virus PCR detection system. The detection system consists of a primer and a probe, Premix Ex Taq reaction solution and sterilized Tris solution, wherein the primer and probe have good detection specificity. The system has high sensitivity, is very suitable for chikungunya virus and does not have cross reactions with the dengue virus and the like.

Description

Technical field [0001] The invention belongs to the field of biological detection, and specifically relates to a rapid qualitative and quantitative detection method for chikungunya virus and a PCR system for chikungunya virus detection, and provides a pair of specific primers and probes. Background technique [0002] Chikungunya fever is an acute natural foci infectious disease caused by Chikungunya virus. It is a togavirus and mosquitoes are the vector. The disease is mainly transmitted by infected animals (African green monkeys, baboons, chimpanzees, cows, horses, pigs, rabbits, etc.) and patients through blood sucking by Aedes aegypti, African mangyes, Aedes africanus, brown-winged mangyes, etc. , Can also spread through the respiratory tract. Chikungunya has occurred many times in Africa and Asia in Indonesia, the Philippines, Thailand, Vietnam, Myanmar and India. Due to the movement of people at home and abroad, chikungunya fever has gradually spread to China. In 1987, a c...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68C12N15/11G01N21/64
CPCY02A50/30
Inventor 杨宇姚李四王静张晓龙孙肖红胡孔新白琳
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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