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LAMP (Loop-mediated Isothermal Amplification) detection reagent of DHBV (Duck Hepatitis B Virus)

A hepatitis B virus, ring-mediated isothermal technology, applied to the determination/testing of microorganisms, methods based on microorganisms, biochemical equipment and methods, etc., can solve the problems of high specificity and sensitivity, difficult operation, and short detection time and other issues to achieve the effect of high specificity and high sensitivity

Pending Publication Date: 2018-11-06
NANYANG NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In order to overcome the defects of low efficiency, difficult operation and high cost of ordinary detection, the purpose of the present invention is to provide a ring-mediated isothermal amplification detection reagent of duck hepatitis B virus. When the reagent is used for detection, the detection time is short and the specificity is high. High sensitivity and sensitivity, easy operation and low cost

Method used

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  • LAMP (Loop-mediated Isothermal Amplification) detection reagent of DHBV (Duck Hepatitis B Virus)
  • LAMP (Loop-mediated Isothermal Amplification) detection reagent of DHBV (Duck Hepatitis B Virus)
  • LAMP (Loop-mediated Isothermal Amplification) detection reagent of DHBV (Duck Hepatitis B Virus)

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Embodiment 1

[0050] 1. Reagent related materials

[0051] Duck hepatitis B virus is a strain isolated and preserved in the laboratory. Viral Genomic DNA / RNA Extraction Kit (Beijing Quanshijin Biotechnology Co., Ltd.), Betaine (SIGMA Company), BstDNA Polymerase and 10×Bst Buffer Reaction Buffer (Biolabs Inc), dNTP (clontech Company), Mg 2+ and DL2000 (TAKARA company). The chromogenic reagent was a mixed staining solution of 0.025 mM phenol red and 0.08 mM cresol red, both of which were purchased from Solarbio.

[0052] 2. Design and synthesis of LAMP primers

[0053] Referring to the currently published duck hepatitis B virus sequence, aiming at the conserved region S region of the DHBV virus genus, 4 primers were designed using the online design software Primer Explorer V4 (F1c and B1c were respectively opposite to the F1 region and B1 region downstream of the amplification. The primers include the upstream internal primer FIP (FIP consists of F1c, F2 and GAATTC (EcoRI restriction site ...

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Abstract

The invention belongs to the technical field of virus detection, and specifically relates to a LAMP (Loop-mediated Isothermal Amplification) detection reagent of DHBV (Duck Hepatitis B Virus). A specific LAMP primer is designed and synthesized according to a DHBV gene conserved region, and the specific LAMP primer comprises an upstream inner primer FIP, a downstream inner primer BIP, an upstream outer primer F3 and a downstream outer primer B3. The LAMP detection reagent is prepared from the following ingredients: a 10*Bst Buffer reaction buffer solution, dNTPs (Deoxynucleotide Triphosphates),MgSO4, Bst DNA (Deoxyribonucleic Acid) polymerase, deionized water, a color-producing reagent and the primers. The LAMP detection reagent provided by the invention has the advantages of strong specificity, high sensitivity, simpleness and convenience in operation, low cost and the like during application.

Description

technical field [0001] The invention belongs to the technical field of virus detection, in particular to a detection reagent for ring-mediated isothermal amplification of duck hepatitis B virus. Background technique [0002] Duck hepatitis B virus (DHBV) and human hepatitis B virus (HBV) belong to the hepadnaviridae family, and they are similar in morphology, gene sequence, virus replication, pathogenic mechanism, and infection host rules. Relatively similar, the DHBV virus particle is spherical, has a capsule, and has a diameter of 40-50nm. It is transmitted vertically through eggs, so ducklings have the highest infection rate and no clinical symptoms after infection. However, it has been reported that its infection can cause liver cell degeneration and focal Hepatitis-like pathological changes such as necrosis and inflammatory cell infiltration in the portal area. The duck model of DHBV infection has enabled people to understand the replication cycle of hepadnavirus and t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11C12R1/93
CPCC12Q1/6844C12Q1/706C12Q2531/119
Inventor 冀君许鑫王雪雨吴倩倩李宛玉王静阚云超姚伦广
Owner NANYANG NORMAL UNIV
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