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Novel gosling gout virus one-step method loop-mediated isothermal detection reagent and application thereof

A loop-mediated isothermal, detection reagent technology, used in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., to achieve high sensitivity and high specificity

Pending Publication Date: 2020-06-19
NANYANG NORMAL UNIV
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Problems solved by technology

[0003] Nucleic acid amplification is one of the most commonly used methods in genetic diagnostic technology. Currently, nucleic acid amplification methods include polymerase chain reaction (PCR), nucleic acid sequence-dependent amplification (NASBA), autonomous sequence replication (SR) and strand displacement techniques. (SDA), etc., can rapidly amplify trace samples, but have their own shortcomings in terms of specificity, simplicity, temperature and reagent instrument requirements.

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  • Novel gosling gout virus one-step method loop-mediated isothermal detection reagent and application thereof
  • Novel gosling gout virus one-step method loop-mediated isothermal detection reagent and application thereof
  • Novel gosling gout virus one-step method loop-mediated isothermal detection reagent and application thereof

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Embodiment Construction

[0051] The following examples facilitate a better understanding of the present invention, but do not limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples are commercially available unless otherwise specified.

[0052] The loop-mediated isothermal amplification technology (loop-mediated isothermal amplification, LAMP) involved in this study can amplify RNA fragments with very low copy numbers within 1 hour under isothermal conditions in a water bath environment. Before the reaction starts, add a pH indicator, and the result can be judged by the naked eye due to the color change after the reaction.

[0053] The characteristic of the loop-mediated isothermal amplification technique is to use 3 pairs of special primers corresponding to the 8 gene segments in the target sequence (outer primers F3, B3; inner primers FIP, BIP; loop primers LF, LB). 3.0 T...

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Abstract

The invention provides a novel gosling gout virus (N-GoAstV) one-step method loop-mediated isothermal detection reagent and application thereof and relates to the technical field of virus detection. According to the detection reagent, specific LAMP (loop-mediated isothermal amplification) primers are designed and synthesized according to an N-GoAstV gene conserved region; the primers comprise an upstream inner primer FIP, a downstream inner primer BIP, an upstream outer primer F3, a downstream outer primer B3, an upstream looped primer LF and a downstream looped primer LB; and the detection reagent comprises the following components: a 10x reaction buffer, dNTPs (deoxynucleotides), MgSO4, Bst 3.0DNA polymerase, deionized water, a developing reagent and the primers. The detection reagent provided by the invention has the advantages of being good in specificity, high in sensitivity, simple and convenient to operate, low in cost, and the like.

Description

technical field [0001] The invention relates to the technical field of virus detection, in particular to a one-step ring-mediated isothermal detection reagent for novel goose gout virus and its application. Background technique [0002] The goose-causing novel Goose Astrovirus (N-GoAstV) is a member of the Astroviridae family. Due to its short discovery time, the genus and species have not yet been determined. Gout-causing novel gosling astrovirus is a non-encapsulated spherical particle with a diameter of 28-30 nm, and its capsid protein has a 20-hedral structure. N-GoAstV infection is widely prevalent in goslings, mainly causing enteritis and nephritis in goslings, with the highest infection rate at 9-12 days old. Due to this age, the level of antibodies from the mother in goslings decreases, coupled with environmental pollution and untimely prevention, once infected, it will cause great damage to the body. After infection, the manifestations are high fever, diarrhea, mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/701C12Q1/6844C12Q2600/166C12Q2531/119C12Q2545/113C12Q2565/125
Inventor 冀君陈钦玺刘强许鑫慕新浩张志斌余政利随朝鸽阚云超姚伦广
Owner NANYANG NORMAL UNIV
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