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Reverse transcription loop-mediated isothermal amplification primer, kit and detection method for visually detecting Hobi-like pest virus

A loop-mediated isothermal and isothermal amplification technology, used in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problem of high operating cost, inability to detect HoBi-like pestivirus, inability to directly Visual observation and other problems, to achieve the effect of high sensitivity, high sensitivity and high specificity

Inactive Publication Date: 2017-05-24
NANYANG NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this patented kit can only detect BVDV-1 strains, and cannot detect HoBi-like pestiviruses. Moreover, it cannot be directly observed with the naked eye by using ultraviolet detection or electrophoresis detection, and the operation cost is high.

Method used

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  • Reverse transcription loop-mediated isothermal amplification primer, kit and detection method for visually detecting Hobi-like pest virus
  • Reverse transcription loop-mediated isothermal amplification primer, kit and detection method for visually detecting Hobi-like pest virus
  • Reverse transcription loop-mediated isothermal amplification primer, kit and detection method for visually detecting Hobi-like pest virus

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Embodiment Construction

[0043] The present invention will be further described below in conjunction with some specific embodiments.

[0044] 1. Kit related materials

[0045] Bovine viral diarrhea virus type 1 (BVDV-1) and bovine herpesvirus type 1 (BHV-1) originated from the China Veterinary Drug Administration; bovine rotavirus (BRV), bovine parainfluenza virus type 3 (BPIV3) and Hobi- Pestivirus like is isolated and preserved in the laboratory. Viral Genomic DNA / RNA Extraction Kit (Beijing Quanshijin Biotechnology Co., Ltd.), Betaine (SIGMA), BstDNA polymerase and 10×Thermo pol reaction buffer (Biolabs Inc), dNTP (clontech), AMV Reverse Transcriptase (promega company), Ribonuclease Ihibitor (NEB company), Mg 2+ And DL2000 (TAKARA company).

[0046] 2. Design and synthesis of LAMP primers

[0047] Referring to the currently published Hobi-like pestivirus sequence (Accession No. GenBank KU563155), for the 5'-UTR region of the conserved region of Pestivirus, relevant primers were designed using t...

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Abstract

The invention belongs to the technical field of virus detection, and particularly relates to a reverse transcription loop-mediated isothermal amplification (LAMP) primer, a reverse transcription LAMP kit and a reverse transcription LAMP detection method for visually detecting a Hobi-like pest virus. An upstream inner primer FIP, a downstream inner primer BIP, an upstream outer primer F3 and a downstream outer primer B3 of the specific LAMP primer are synthesized; the kit is prepared from the following components: a 10*Thermo Pol reaction buffer solution, dNTP, MgSO4, Bst DNA polymerase, AMV Reverse Transcriptase, a Ribonuclease Ihibitor, DEPC water, a developing reagent and the primer. The kit is high in specificity, high in sensitivity, quick and high in operability.

Description

technical field [0001] The invention belongs to the technical field of virus detection, and in particular relates to a reverse transcription loop-mediated isothermal amplification primer, a kit and a detection method for visually detecting Hobi-like pestiviruses. Background technique [0002] Hobi-like pestivirus is a new pestivirus isolated from Brazilian fetal bovine serum in 2004, which is related to bovine viral diarrhea virus type 1 (BVDV-1), bovine viral diarrhea virus type 2 (BVDV-2), Both swine fever virus (CSFV) and sheep border virus (BDV) belong to the family Flaviviridae. Later, the distribution and existence of the virus were found in other countries, such as Italy and Switzerland in Europe, Australia in Oceania, Mexico in the Americas, Japan, India and Bangladesh in Asia. The symptoms caused by the virus after infecting cattle are similar to those shown by BVDV-1 and BVDV-2 infected cattle. The main clinical manifestations are respiratory tract, digestive trac...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6844C12Q1/701C12Q2531/119C12Q2521/107C12Q2545/113
Inventor 史鸿飞阚云超姚伦广唐存多焦铸锦冷超粮冀君岳超马娜
Owner NANYANG NORMAL UNIV
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