Fluorescence quantitative polymerase chain reaction (PCR) new method for detecting multiple viruses of yellow fever, dengue fever and epidemicencephalitis B and multiple virus detection PCR system

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

Active Publication Date: 2011-09-21
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Abstract
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Problems solved by technology

[0005] All three viruses belong to flaviviruses. Currently, methods for detecting flaviviruses include reverse transcription polymerase reaction, SYBR Green Real-time PCR, microwell hybridization, and protein chip technology. Simultaneous detection of flaviviruses, and this method is undoubtedly faster and more accurate

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  • Fluorescence quantitative polymerase chain reaction (PCR) new method for detecting multiple viruses of yellow fever, dengue fever and epidemicencephalitis B and multiple virus detection PCR system
  • Fluorescence quantitative polymerase chain reaction (PCR) new method for detecting multiple viruses of yellow fever, dengue fever and epidemicencephalitis B and multiple virus detection PCR system
  • Fluorescence quantitative polymerase chain reaction (PCR) new method for detecting multiple viruses of yellow fever, dengue fever and epidemicencephalitis B and multiple virus detection PCR system

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Embodiment Construction

[0048] The working principle of the new method for fluorescent quantitative PCR detection of yellow fever, dengue, and Japanese encephalitis multiple viruses is to use the qualitative analysis of the change of the fluorescent signal to detect the change of the amount of each cycle of amplification product in the PCR amplification reaction in real time, The starting template was quantified by the relationship between the Ct value and the standard curve. The present invention also relates to all the contents contained in the above-mentioned detection system.

[0049] The method includes the following steps:

[0050] 1) Design primer probes;

[0051] 2) Select fluorescein according to the instrument;

[0052] 3) Synthetic primers and probes;

[0053] 4) Prepare positive plasmid standard;

[0054] 5) run PCR;

[0055] 6) Data analysis;

[0056] 7) Extract viral RNA for system verification.

[0057] 1. Design of primers and probes

[0058] According to the names of the gene...

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Abstract

The invention discloses a fluorescence quantitative polymerase chain reaction (PCR) new method for detecting multiple viruses of yellow fever, dengue fever and epidemicencephalitis B and multiple virus detection PCR system consisting of primers, probes, a Premix Ex Taq reaction solution and a sterilized Tris buffer. With good singularity and high sensitivity, three pairs of primers and probes are very suitable for simultaneously detecting viruses of yellow fever, dengue fever and epidemicencephalitis B. And there is no cross reaction between the primers and probes and several other entomophily hemorrhagic fever viruses, such as Marburg virus and Rift Valley fever virus.

Description

technical field [0001] The invention belongs to the field of biological detection, in particular to rapid qualitative and quantitative detection methods for yellow fever, dengue and Japanese encephalitis viruses, each of which provides a pair of specific primers and probes. Background technique [0002] Yellow fever (commonly known as "yellow jack", "black vomit", and sometimes the American plague) is an acute viral disease. It has caused some devastating diseases. In the transmission of the disease from monkeys to humans or between humans, mosquitoes are the main mediators. Every year there are at least 200,000 cases of yellow fever worldwide, resulting in 30,000 deaths, 90% of which are from Africa. Due to the high mortality rate and strong contagiousness of yellow fever, it has been included as one of the quarantine infectious diseases stipulated by the World Health Organization. [0003] Epidemic encephalitis (epidemicencephalitisB) is a central nervous system infecti...

Claims

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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 杨宇白琳王静胡健萍马雪征杨志红
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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