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Primers and probes for the detection of foot-and-mouth disease virus universal, type a duplex real-time fluorescent quantitative PCR

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

Active Publication Date: 2016-06-29
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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  • Primers and probes for the detection of foot-and-mouth disease virus universal, type a duplex real-time fluorescent quantitative PCR
  • Primers and probes for the detection of foot-and-mouth disease virus universal, type a duplex real-time fluorescent quantitative PCR
  • Primers and probes for the detection of foot-and-mouth disease virus universal, type a duplex real-time fluorescent quantitative PCR

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0032] 1. Materials and methods

[0033] 1.1 Material

[0034] 1.1.1 Strain

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

[0036] 1.1.2 Instruments and reagents

[0037] Fluorescence PCR instrument, American ABI company product, model ABI7000; PCR amplification instrument, German Biometra company product; Gel imaging analysis system, American AlphaInnotech product; Constant temperature water bath oscillator (HZQ-Q), Harbin Donglian Electronic Technology Development Co., Ltd. Company products; desktop high-speed refrigerated centrifuge, American Heraeus products; ExTaq DNA polymera...

Embodiment 2

[0070] Example 2 Application of foot-and-mouth disease virus universal, type A double real-time fluorescent quantitative PCR detection system

[0071] According to the FMDV-T / FMDV-A dual FQ-PCR established in this research and the FMDV-T / FMDV-A common RT-PCR detection method established by our laboratory, the detection reagents are prepared to FMDV-T / FMDV-A The positive plasmid mixture is the positive control, and the water is the negative control. 13 clinically suspected FMDV infection samples (sample numbers: 1-13) were tested for application. First, the 13 samples and the negative and positive controls were homogenized by a homogenizer. Centrifuge at 5000r / min for 5 minutes, take 100μL of the supernatant, extract total RNA with a nucleic acid extractor, use the method provided by the present invention, foot-and-mouth disease virus O, A and Asia I triple RT-PCR detection reagents and detection methods (ordinary RT -PCR test, authorized announcement number CN102230029B) Laborato...

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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 specifically relates to foot-and-mouth disease virus universal and type A double real-time fluorescent quantitative PCR detection primers and probes. Background technique [0002] Foot-and-mouth disease (FMD) is an acute, malignant and highly contagious infectious disease caused by foot and mouth disease virus (Foot and Mouth Disease Virus, FMDV), which is caused by pigs, cattle, sheep and other major domestic animals and other domestic and wild cloven-hoofed animals. Many kinds. The disease spreads in many ways and fast, and it has repeatedly spread worldwide, causing huge political and economic losses. The World Organization for Animal Health (OIE) lists it as the first category A infectious disease. At present, FMD is prevalent in two-thirds of OIE member countries, which is a constant threat to livestock safety and livestock product trade in FMD-free countries...

Claims

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Application Information

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Patent Type & Authority Patents(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
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