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Method and kit for detecting reverse transcription-loop mediated isothermal amplification (RT-LAMP) of pike fry rhabdovirus (RFRV)

A technology for constant temperature amplification of detection kits and ring media, which is applied to biochemical equipment and methods, and microbial measurement/inspection. It can solve the problems of long cycle and inaccurate detection results, and achieve short time consumption, high sensitivity, good specific effect

Inactive Publication Date: 2013-06-12
SHENZHEN AUDAQUE DATA TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This detection method not only has a long cycle, but also uses immunological methods, such as indirect fluorescent antibody test or enzyme-linked immunosorbent assay, there is a cross-reaction between PFRV and SVCV ( et al., 1989; Way, 1991), the test results are not accurate

Method used

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  • Method and kit for detecting reverse transcription-loop mediated isothermal amplification (RT-LAMP) of pike fry rhabdovirus (RFRV)
  • Method and kit for detecting reverse transcription-loop mediated isothermal amplification (RT-LAMP) of pike fry rhabdovirus (RFRV)
  • Method and kit for detecting reverse transcription-loop mediated isothermal amplification (RT-LAMP) of pike fry rhabdovirus (RFRV)

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Embodiment 1

[0063] 1 Materials and methods

[0064] 1.1 Viruses and cells

[0065] PFRV is the F4 strain, donated by the Institute of Hydrobiology, Chinese Academy of Sciences, SVCV and IHNV reference strains were donated by Prof. B. Hill, OIE Reference Laboratory of CEFAS, UK; VHSV reference strain was donated by Dr. Roar Gudding, OIE Reference Laboratory, Norwegian National Veterinary Research Institute , IPNV, HRV, and VNNV are separated by this room.

[0066] IHNV, SVCV, VHSV, VNNV, IPNV, HRV and PFRV were amplified with EPC, CO, RTG-2, SSN-1, CHSE-214, FHM and BF-2 cells, respectively. SSN-1 cells were purchased from the Institute of Aquatic Animal Health, Kasetsart University, Thailand, RTG-2 cells were donated by the Institute of Hydrobiology, BF-2 cells were donated by the Cell Collection Center of the Institute of Quarantine (Brescia), CHSE-214 cells, EPC cells were donated by The British Environment, Fisheries and Aquaculture Science Research Center donated, the FHM cells were...

Embodiment 2

[0117] A detection kit for pike juvenile rhabdovirus (Pike fry rhabdovirus, PFRV), the kit includes the following two pairs of primers:

[0118] Primer

[0119] The inner primers FIP and BIP contain two different sequences, F1c+F2 and B1c+TTTT+B2, respectively. The kit also includes instructions for use. This kit can be used for RT-LAMP detection of rhabdovirus in juvenile pike.

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Abstract

The invention discloses a method and a kit for detecting reverse transcription-loop mediated isothermal amplification (RT-LAMP) of pike fry rhabdovirus (RFRV), wherein an adopted outer primer contains sequences indicated by Seq ID No.1 and Seq ID No.2 respectively. According to the method and the kit which are provided by the invention, PFRV can be detected specifically; and the sensitivity is high and particularly reaches 100.5TCID50 when the kit is used for detecting the extracted nucleic acid in a virus suspension.

Description

technical field [0001] The invention relates to a virus detection method and a detection kit, in particular to a detection method and a kit for juvenile pike rhabdovirus reverse transcription cyclase constant temperature amplification detection. Background technique [0002] According to the 8th report of the International Committee on Taxonomy of Viruses (ICTV), Rhabdoviridae (Rhabdoviridae) is divided into 6 genera, including the plastotype Rhabdoviruses that infect plants ( Cytorhabdovirus ) and nuclear rhabdoviruses ( Nucleorhabdovirus ), and vesicular stomatitis viruses that infect vertebrates ( Vesiculovirus ), lyssavirus ( Lyssavirus ), Ephemerovirus and extragranular rhabdovirus (also known as non-virulent rhabdovirus) ( Novirhabdovirus ), and some rhabdoviruses have not yet been classified, such as Sigma virus, Flanders virus, etc. So far, more than a dozen rhabdoviruses have been reported to infect fish, including infectious hematopoietic necrosis virus (IH...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
Inventor 刘荭史秀杰卢体康陈红莲何俊强
Owner SHENZHEN AUDAQUE DATA TECH
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