Reagent, detection method and application for detection of African hog cholera virus

A technology of African swine fever virus and reagents, which is applied in the field of detection of African swine fever virus, can solve the problems of complicated transmission of the epidemic and the complicated and severe situation of prevention and control, and achieve the effects of preventing the spread of the epidemic, fast detection speed, and simple and convenient operation

Inactive Publication Date: 2019-05-10
SHENZHEN AUDAQUE DATA TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But at the same time, the traditional breeding structure is difficult to fundamentally change in a short period of time, the transmission of the epidemic is complicated, and the prevention and control situation is still complicated and severe. Fast and efficient on-site detection is of great significance to the prevention and control of African swine fever

Method used

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  • Reagent, detection method and application for detection of African hog cholera virus
  • Reagent, detection method and application for detection of African hog cholera virus
  • Reagent, detection method and application for detection of African hog cholera virus

Examples

Experimental program
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Effect test

Embodiment

[0039] 1. Materials and methods

[0040] 1. Test nucleic acid

[0041] In this example, according to the main structural protein gene B646L gene (p72 gene) of African swine fever virus published in NCBI GenBank, the gene fragment was synthesized by Shenggong Bioengineering (Shanghai) Co., Ltd. and cloned into the pMD19-T vector, named ASFV -p72. Other nucleic acid templates: swine fever virus (CSFV), porcine reproductive and respiratory syndrome (PRRSV), porcine circovirus type II (PCV2), foot-and-mouth disease virus (FMDV), porcine Seneca virus type A (SVA), etc. Inactivate viral nucleic acid.

[0042] 2. Main reagents and instruments

[0043] TwistAmp exo kit is a product of TwistDx Company in the UK, BioDrop ultra-trace nucleic acid and protein concentration analyzer, Axxin constant temperature amplification instrument, Eppendorf5415R high-speed centrifuge, etc.

[0044] 3. Design and screening of RPA primers and probes

[0045] In this case, a number of specific prime...

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Abstract

The invention discloses a reagent, detection method and application for the detection of African hog cholera virus. The reagent comprises a specific primer pair and a probe, upstream and downstream primers of the primer pair are respectively sequences shown in Seq ID No. 1 and 2, and the probe is a sequence shown in Seq ID No. 3 or its reverse complement sequence; in the probe sequence shown in the SEQ ID No.3, a 28th base modifies fluorescent quenching group-dT, a 31st base is replaced with a base analog, a 33rd base modifies a fluorescent group-dT, and a 3' end modifies C3 Spacer. The reagent is sensitive, specific and efficient for detection of the African hog cholera virus by recombinase polymerase amplification; compared with conventional conventional or real-time fluorescent PCR, thereagent and the detection method have short detection time and simple operation, are especially suitable for on-site testing, and are of great significance for the rapid prevention and control of theAfrican hog cholera virus and guarantee of production safety.

Description

technical field [0001] The application relates to the field of detection of African swine fever virus, in particular to a reagent, detection method and application for detection of African swine fever virus. Background technique [0002] African swine fever (African swine fever, ASF) is an acute, virulent and highly contagious disease of pigs caused by African swine fever virus (ASF virus, ASFV), characterized by high fever, reticuloendothelial system hemorrhage and high mortality as a feature. Infection with super-virulent strains of ASFV can cause 100% death of pigs within 12-14 days, and the mortality rate of pigs infected with moderate-virulence strains is generally 30%-50%, which is extremely harmful to the pig industry. The World Organization for Animal Health (OIE) lists African swine fever as an important animal infectious disease, and the Ministry of Agriculture and Rural Affairs of my country lists it as a first-class animal disease. [0003] ASFV is the only mem...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12R1/93
Inventor 花群义林彦星花群俊曾少灵曹琛福杨俊兴
Owner SHENZHEN AUDAQUE DATA TECH
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