Primer and kit for efficiently detecting Erysipelothrix rhusiopathiae

A porcine erysipelas and kit technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., can solve the problems of insufficient specificity, high cost, and many steps in micro-dosing, so as to avoid non-specific The effect of heterosexual reaction and easy operation

Pending Publication Date: 2020-12-04
宁波爱基因科技有限公司
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AI Technical Summary

Problems solved by technology

If the detection of this virus adopts the existing PCR technology, because its reaction needs to cycle in two different temperature zones, the requirements for the instrument are high, and the cost is relatively high, and the PCR technology only has one pair of amplification primers, which is susceptible to interference and specificity. Relatively insufficient performance, and the time to produce results is long, the operation requires high professionalism, and there are many steps for micro-dosing
The LAMP technology has 4 different specific primers, so the accuracy of the detection results is higher, but because it is a constant temperature reaction, it lacks a hot-start enzyme similar to PCR, and it is easy to produce non-specific amplification during the heating stage of the equipment, which will affect the detection results.

Method used

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  • Primer and kit for efficiently detecting Erysipelothrix rhusiopathiae
  • Primer and kit for efficiently detecting Erysipelothrix rhusiopathiae
  • Primer and kit for efficiently detecting Erysipelothrix rhusiopathiae

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Embodiment 1

[0028] First search for the target sequence through NCBI GenBank, and design primers for the target sequence, and fix them on the corresponding positions of the microfluidic chip, then package the microfluidic chip, and use pork and its various organs, blood, excrement , environmental samples and nucleic acid templates extracted from cell culture are mixed and reacted, then added to the packaged microfluidic chip, and then put into the microfluidic chip detector with centrifugal function, constant temperature function and real-time fluorescence detection , the above-mentioned instruments and chips are all commercially available products, and Ningbo Ai Gene Technology Co., Ltd. also has corresponding public products, which use centrifugal force to drive samples into the microfluidic chip reaction wells for constant temperature amplification. If the sample contains the target fragment and is amplified at constant temperature, the amplified product is effectively combined with the...

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Abstract

The invention discloses a primer and kit for efficiently detecting Erysipelothrix rhusiopathiae. According to the primer and the kit, an improved LAMP technology serves as a gene amplification reaction principle, and gold nanoparticles are added into a reaction system to adsorb ssDNA and protease and inhibit nonspecific reactions during heating, thus, the aim of warm start is achieved, and the nonspecific reactions during heating are avoided; by combining the improved LAMP technology and a microfluidic chip technology, an accurate detection result can be presented rapidly, and the aim of jointdetection on the same sample with a plurality of different indexes is achieved; and meanwhile, reaction reagents are pre-buried into a microfluidic chip, a user only needs to add a sample, and operations are simple and convenient.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a primer and a kit for efficiently detecting Erysipelothrix suis. Background technique [0002] Swine erysipelas (SER), also known as "fire print", is a second-class infectious disease and parasitic disease. It is an acute, febrile zoonotic infectious disease caused by E. rhusiopathiae, which belongs to the second category of animal diseases announced by the Ministry of Agriculture. The disease widely exists in all parts of the world, mainly occurs in pigs, and it is most likely to attack sows and shelf pigs. The initial stage of the disease is mostly acute septicemia or subacute rash type, and then it turns into chronic type. The affected pigs often have arthritis, Endocarditis. If the existing PCR technology is used for the detection of the virus, since the reaction needs to be circulated in two different temperature zones, the requirements for the instrume...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/6844C12Q1/04C12N15/11
CPCC12Q1/689C12Q1/6844C12Q2600/166C12Q2531/119C12Q2563/107C12Q2563/137C12Q2565/629C12Q2545/113
Inventor 卢先东刘艳红乙丛敏刘萌
Owner 宁波爱基因科技有限公司
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