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Test strip kit for detecting nucleic acid of classical swine fever virus through room-temperature recombinase polymerase amplification (RT-RPA) and application thereof

A nucleic acid test strip and swine fever virus technology, which is used in the field of rapid detection of swine fever virus, can solve problems such as the lack of primer probe design software, and achieves fast detection speed, simple operation and good specificity. Effect

Pending Publication Date: 2017-12-05
SOUTH CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

RPA technology is in the initial research stage, there is no special primer and probe design software, and there is no large amount of data to provide a basis for its primer design principles

Method used

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  • Test strip kit for detecting nucleic acid of classical swine fever virus through room-temperature recombinase polymerase amplification (RT-RPA) and application thereof
  • Test strip kit for detecting nucleic acid of classical swine fever virus through room-temperature recombinase polymerase amplification (RT-RPA) and application thereof
  • Test strip kit for detecting nucleic acid of classical swine fever virus through room-temperature recombinase polymerase amplification (RT-RPA) and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Establishment of the RT-RPA nucleic acid test strip kit and detection method for rapid detection of classical swine fever virus.

[0056] 1. Design and preparation of primers and probe sequences

[0057] The inventors of the present invention have passed AY259122.1, NC_002657.1, AY382481.1, HQ380231.1, KP233071.1, KM3622426.1, AY775178.2, KT119352.1, X87939.1, AF326963.1 from Genbank , KT716271.1, and EU915211.1 NS5B gene homologous sequences were compared and analyzed, and the conserved region of the NS5B gene of CSFV was determined. According to the RPA primer design principle, primers and probes were designed for this region in order to detect As much swine fever virus as possible. The requirement of RPA for primer length is 30-35bp, the amplification product is within 500bp, and the amplification efficiency is high. To establish a fast and sensitive detection method, it is necessary to screen multiple primers, and individual base differences in the prime...

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Abstract

The invention discloses a test strip kit for detecting nucleic acid of classical swine fever virus through room-temperature recombinase polymerase amplification (RT-RPA) and application thereof. The kit comprises primer pairs with nucleotide sequence as shown in the claim 1 and a nucleic acid test strip. An application method of the kit comprises the following steps: firstly a RT-RPA reaction system is configured for normal-temperature constant-temperature reaction; after detected with the nucleic acid test strip, a product obtained after the constant-temperature reaction directly undergoes interpretation: a positive result is two red strips with one strip positioned in a detection area and the other strip positioned in a quality control area; a negative result is a red strip in the quality control area with no red strip in the detection area. The RPA-nucleic acid test strip technology is adopted for the first time to establish a method for rapid detection of CSFV. The kit of the invention is simple to operate and has high specificity. Reaction results are easy to observe. The kit is safe and pollution-free, and is very suitable for clinic field test.

Description

technical field [0001] The present invention relates to a kind of test kit for detecting swine fever virus and its application, in particular to a test strip RPA kit for rapid detection of swine fever virus based on RPA reaction, and also to its use in the rapid detection of swine fever virus , The invention belongs to the field of preventive veterinary inspection. Background technique [0002] Classical swine fever (CSF) is a major and severe infectious disease of pigs that endangers the development of the breeding industry. It is mainly caused by the infection of classical swine fever virus (CSFV). Reduction is the main feature, and it is one of the communicable diseases that must be notified in the list of epidemic diseases listed by the World Health Organization (OIE). After pigs are infected with CSFV, depending on the virulence of the strains and the age and breed of the infected pigs, the disease can be manifested in various forms such as mild or severe, which mainly...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/701C12Q1/6844C12Q2521/507C12Q2521/101C12Q2565/625
Inventor 陈金顶张静远范双旗徐海娈刘志香赵明秋李军
Owner SOUTH CHINA AGRI UNIV
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