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General A-type double real-time fluorescence quantitation PCR detection primer and probe for foot and mouth disease viruses

A real-time fluorescent quantitative, foot-and-mouth disease virus technology, applied in the field of animal disease quarantine and detection, which can solve the problems of political, economic losses, and multiple transmission routes.

Active Publication Date: 2015-03-11
HENAN CENT FOR ANIMAL DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disease has many transmission routes and fast speed, and has repeatedly broken out in the world, causing huge political and economic losses.

Method used

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  • General A-type double real-time fluorescence quantitation PCR detection primer and probe for foot and mouth disease viruses
  • General A-type double real-time fluorescence quantitation PCR detection primer and probe for foot and mouth disease viruses
  • General A-type double real-time fluorescence quantitation PCR detection primer and probe for foot and mouth disease viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 The establishment of a general-purpose, A-type double real-time fluorescent quantitative PCR detection system for foot-and-mouth disease virus

[0032] 1. Materials and methods

[0033] 1.1 Materials

[0034] 1.1.1 Strains

[0035] The standard strains of type O, type A and Asia I foot-and-mouth disease virus were purchased from the Lanzhou Veterinary Research Institute of the Ministry of Agriculture; other control viruses or plasmids were preserved by the Animal Disease Prevention and Control Center of Henan Province.

[0036] 1.1.2 Instruments and reagents

[0037] Fluorescence PCR instrument, product of ABI Company in the United States, model ABI7000; PCR amplification instrument, product of Biometra Company in Germany; gel imaging analysis system, product of Alpha Innotech Company in the United States; constant temperature water bath oscillator (HZQ-Q), developed by Harbin Donglian Electronic Technology Co., Ltd. products; desktop high-speed refrigerate...

Embodiment 2

[0070] Embodiment 2 The application of foot-and-mouth disease virus universal, type A dual real-time fluorescent quantitative PCR detection system

[0071] According to the FMDV-T / FMDV-A double FQ-PCR established in this study and the FMDV-T / FMDV-A common RT-PCR detection method established in this laboratory, the detection reagents were prepared, and the FMDV-T / FMDV-A The positive plasmid mixture is the positive control, and the water is the negative control. 13 samples of clinically suspected FMDV infection (sample number: 1-13) were tested. First, the 13 samples and the negative and positive controls were homogenized by homogenizer , 5000r / min centrifugal 5 minutes, get supernatant 100 μ L, nucleic acid extractor extracts total RNA, utilize method provided by the present invention and foot-and-mouth disease virus O type, A type and Asia I triple RT-PCR detection reagent and detection method thereof (common RT -PCR detection, authorized announcement number CN 102230029B) Lab...

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Abstract

The invention provides a general A-type double real-time fluorescence quantitation PCR (Polymerase Chain Reaction) detection primer and probe for foot and mouth disease viruses. The primer comprises P0, P1, P2, P3 and P4, and the probe comprises P5 and P6. Double real-time fluorescence quantitation PCR detection established based on the primer and the probe can realize quick identification and detection for three subtype general foot and mouth disease viruses, namely the foot and mouth disease viruses O, A, AsinaI in one reaction at the same time, and the detection can be completed in 1-2 h; the detection has the advantages of being quick, specific, sensitive, high in flux and the like, and the requirements for quickly identifying and detecting the foot and mouth disease viruses and A-type foot and mouth disease viruses in a large scale can be met.

Description

technical field [0001] The invention belongs to the technical field of animal disease quarantine and detection, and in particular relates to primers and probes for dual real-time fluorescent quantitative PCR detection of foot-and-mouth disease virus general type and type A. Background technique [0002] Foot-and-mouth disease (FMD) is an acute, vicious, highly contagious infectious disease caused by foot-and-mouth disease virus (FMDV) caused by pigs, cattle, sheep and other main livestock and other domestic and wild cloven-hoofed animals. There are more than 70 kinds of animals. The disease has many transmission routes and fast speed, and has repeatedly broken out in the world, causing huge political and economic losses. The World Organization for Animal Health (OIE) ranks it as the first class A infectious disease. At present, FMD is prevalent in two-thirds of OIE member countries, threatening the safety of livestock and the trade of livestock products in non-FMD countrie...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/6851C12Q1/701C12Q2600/112C12Q2561/101C12Q2537/143
Inventor 吴志明闫若潜谢彩华赵雪丽陈涛赵明军郑岩严平王淑娟刘琨曹伟伟张代宝王翠韩庆斌朱凤霞王永强
Owner HENAN CENT FOR ANIMAL DISEASE CONTROL & PREVENTION