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Brucella recombinase polymerase amplification detection kit and application thereof

A technology of recombinase polymerase and Brucella, which is applied in the field of genetic engineering technology and diagnostic reagents, can solve problems such as unfavorable detection, and achieve the effect of rapid detection, good sensitivity and simple operation process

Pending Publication Date: 2019-03-19
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Quantitative PCR detection is currently an effective detection method with high specificity, but it is not conducive to field detection due to the need for PCR instruments and the limitations of personnel in the operating site

Method used

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  • Brucella recombinase polymerase amplification detection kit and application thereof
  • Brucella recombinase polymerase amplification detection kit and application thereof
  • Brucella recombinase polymerase amplification detection kit and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0041] 1 Experimental process

[0042] 1.1 Experimental reagents and instruments

[0043] Reagents: RPA Primer Probe, Rehydration Buffer, RNase Inhibitor and dH 2 O, magnesium acetate, 5×Extration Buffer, TaKaRa MiNiBEST Universal Genomic DNA Extraction KitVer.5.0. Lateral flow test paper was purchased from ABINGDON HEALTH company, and its trade name was Nucleic Acid Detection-Nucleic Acid Detection-PCRD.

[0044] Instruments: constant temperature amplification instrument, single-channel pipette, vortexer, small centrifuge.

[0045] 1.2 Genome Extraction

[0046] Genome extraction refers to the TaKaRa MiNiBEST Universal Genomic DNA Extraction KitVer.5.0 reagent manual for extraction. The required detection samples and the genomes of other bacteria in the specificity experiment are all based on this method. The specific method is as follows:

[0047] 1.①Take 10 mg of tissue for grinding, add 200 μl of PBS solution or RnasefreedH to the ground tissue 2 O.

[0048] ② Add 20...

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Abstract

The invention belongs to the technical fields of genetic engineering and diagnostic reagents and particularly relates to a brucella recombinase polymerase amplification (RPA) detection kit and application thereof. The detection kit comprises a specific primer group and a probe. An RPA amplification product is detected by virtue of a test strip and is dropwise added to the test strip after being amplified for only about 10 minutes, and a result is read, so that a PCR amplification instrument and a quantitative PCR instrument are saved. A method provided by the invention has the advantages of strong specificity, good sensitivity, simple operation process, rapidness in detection and the like, the possibility is provided for the detection of field brucellosis, and a method which does not aimsat the diagnosis and treatment of diseases is provided for the research of brucella.

Description

technical field [0001] The invention belongs to the technical fields of genetic engineering technology and diagnostic reagents, and in particular relates to a Brucella recombinase polymerase amplification (RPA) detection kit and its application. Background technique [0002] Brucellosis (brucellosis), referred to as brucellosis, is a bacterial zoonotic disease caused by Brucella, which has acute, subacute and chronic forms. The pathogen Brucella is a Gram-negative facultative intracellular parasite, which can infect susceptible animals through the mouth, nose, throat, conjunctiva, etc., causing the main symptoms such as abortion, stillbirth, and orchitis in male animals, accompanied by Symptoms such as fever, joint pain, and swollen lymph nodes have caused huge economic losses to the development of the aquaculture industry. At the same time, as a typical zoonotic disease, it also poses a great threat to human health. [0003] Currently, the main diagnostic techniques for la...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/04C12N15/11
CPCC12Q1/689
Inventor 曹小安周建华刘永生尚佑军李学瑞兰喜葛志毅
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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