Kit for diagnosing reovirus genes of grass carps and application thereof

Abstract

The invention relates to a kit for diagnosing the reovirus genes of grass carps, comprising an RNA (Ribonucleic Acid) extraction reagent for viruses in visceral tissues, an RAN extraction reagent for cell infecting viruses, an RT (Reverse Transcriptase) reaction reagent and a PCR (Polymerase Chain Reaction) reaction reagent, wherein the RNA extraction reagent for the viruses in the visceral tissues comprises the components of DEPC (Diethyl Pyrocarbonate) water, protease K (1 mg / mL, pH is 8.0), SDS (Sodium Dodecyl Sulfate) (1%), phenol / chloroform / isoamylol (25 / 24 / 1), chloroform / isoamylol (24 / 1), isopropanol and 70% ethanol; the RAN extraction reagent for the cell infecting viruses comprises the components of Trizol, chloroform, isopropanol, 70% ethanol and the DEPC water; the RT reaction reagent comprises the components of 5*AMV Buffer, dNTP, the DEPC water, a reverse primer R1, an RNA enzyme inhibitor RNAsin (R1) and a reverse transcriptase AMV; and the PCR reaction reagent comprises the components of 10*PCR buffer (containing Mg<2+>) and DNTP Mixture (which are respectively 2.5 mmol / L), a specific oligonucleotide primer F1, the primer R1, the DEPC water and Taq E (5U / muL), the forward primer F1:5'-ATCCCGTATATCTATGGCTT-3', and the reverse primer R1:5'TTGGAGACGAACATAGACGC-3. The kit is used for diagnosing the reovirus genes of the grass carps.

Description

technical field [0001] The diagnostic kit and application of the aquatic animal virus of the present invention are mainly a diagnostic kit and a detection method for the conserved part of the sixth gene segment of Grass carp reovirus (GCRV, Grass carp reovirus). Background technique [0002] Grass Carp Hemorrhage Virus (GCHV) belongs to aquatic reovirus, the virus particles are spherical particles with icosahedral symmetry, the diameter of the mature virus is 75nm, it has a double-layer capsid, no envelope, and its genome Consisting of 11 dsRNAs, it is very stable in the range of pH3-11 and resistant to chloroform and ether. The fifth meeting of the International Committee on Taxonomy of Viruses decided to formally create a new genus of aquatic reovirus (Aquareovirus, ARV) in the family Reoviridae, and to include grass carp hemorrhagic disease virus into this genus, named Grass carp reovirus (Grass carp reovirus). reovirus, GCRV), but it is different from the representative...

AI Technical Summary

Problems solved by technology

Although the GCRV RT-PCR method has also been reported in the literature, only the original strain virus used to design primers is PCR positive, and the corresponding fragments of other GCRVs cannot be amplified. This test is based on the GCRV and other aquatic reoviruses published by GenBank. A pair of GCRV-specific primers were designed in the conserved region, and a rapid diagnosis method for GCRV was established, which has not been reported at home and abroad.

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