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Kit for testing three kinds of viruses of fish diseases synchronistically and test method thereof

A technology of simultaneous detection and kits, applied in biochemical equipment and methods, microbial measurement/testing, etc., can solve the problems of complex and cumbersome operation steps and long time, and achieve good specificity, accurate methods, and high practical value Effect

Inactive Publication Date: 2008-01-30
LIANYUN PORT IMMIGRATION INSPECTION & QUARANTINE BUREAU PEOPLES REPUBLIC OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no kits for the detection of these three fish diseases. Currently, the commonly used detection method for the three fish diseases is to inoculate sensitive fish and cells with disease materials for virus propagation, isolate and purify the virus, and then use serum neutralization, immunofluorescence and Enzyme-linked immunoassay method for virus identification, the operation steps are complicated and cumbersome, and the time is as long as 2-4 weeks

Method used

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  • Kit for testing three kinds of viruses of fish diseases synchronistically and test method thereof
  • Kit for testing three kinds of viruses of fish diseases synchronistically and test method thereof
  • Kit for testing three kinds of viruses of fish diseases synchronistically and test method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1. A kit for synchronous detection of fish infectious hematopoietic necrosis virus, infectious pancreatic necrosis virus and viral hemorrhagic sepsis virus, which consists of the following items,

[0060] (1) Negative control, without IHNV, IPNV and VHSV 3 kinds of viral nucleic acid components

[0061] Tissue extract, 1 stick;

[0062] (2) Positive control, containing IHNV, IPNV and VHSV 3 kinds of viral nucleic acid components

[0063] Solution, 1 stick;

[0064] (3) dNTP, 1 branch;

[0065] (4) Contains Mg 2+ PCR buffer, 1 stick;

[0066] (5) Primer pair ihnf / ihnr, 1 each;

[0067] ihnf is: 5'-gttaacttcaacgccaacagg,

[0068] ihnr is: 5'-tgaagtacccctccccgagcatcc;

[0069] (6) Primer pair ipnf / ipnr, 1 each;

[0070] ipnf is: 5'-ccgcaacttacttgagatccattatgc,

[0071] ipnr is: 5'-cgtctggttcagattccacctgtagtg;

[0072] (7) Primer pair vhsf / vhsr, 1 each;

[0073] vhsf is: 5'-cgaccagctcaactcaggtgtcc,

[0074] vhsr is: 5'-cc...

Embodiment 2

[0080] Example 2. The test kit in embodiment 1, it also contains following items:

[0081] (1) Trizol solution for RNA extraction, 1 tube;

[0082] (2) Chloroform, 1 stick;

[0083] (3) Isoamyl alcohol, 1 stick;

[0084] (4) 70% ethanol, 1 stick;

[0085] (5) Aqueous solution containing 0.01% DEPC, 1 bottle.

Embodiment 3

[0086] Example 3. Refer to attached picture. A detection method for simultaneous detection of fish infectious hematopoietic necrosis virus, infectious pancreatic necrosis virus and viral hemorrhagic sepsis virus, using the kit described in Example 1, and the remaining reagents are prepared by the tester , the steps are as follows,

[0087] (1) Extraction of RNA template in the sample: Take 100 μl of susceptible tissue sample homogenate or suspected virus infection suspension, add 500 μl Trizol reagent, vortex for 30 s, add 200 μl chloroform and isoamyl alcohol mixture prepared at a ratio of 24:1, Vortex for 30s, centrifuge at 10000r / min for 5min, take the supernatant, add an equal amount of isopropanol solution and centrifuge as above, take the precipitate, add 300μl of 70% ethanol solution and centrifuge as above, remove the supernatant, air dry, add 50μl of 0.01% DEPC aqueous solution , take 2μl for RT-PCR;

[0088] (2) Preparation of RT-PCR premix: add each reagent in ad...

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Abstract

This invention provides an apparatus which simultaneously detects the fish virus of IHNV, IPNV, and VHSV. It also discloses the method of detecting by this apparatus of solution box. It character in that it's fast and simple with high sensitivity to detect the three bacteriums qualitative. This invention can be used to detect three kinds of disease of import and export, and to monitor epidemic situation of breeding farm to avoid disease dissemination, so that it has a high value.

Description

technical field [0001] The invention relates to a detection kit for aquatic animals, especially fish viruses, in particular to a kit for synchronous detection of fish infectious hematopoietic organ necrosis virus, infectious pancreatic necrosis virus and viral hemorrhagic sepsis virus , the present invention also relates to the aforementioned detection method for synchronous detection. Background technique [0002] Viral hemorrhagic septicemia virus (VHSV), infectious pancreatic necrosis virus (IPNV) and fish infectious hematopoietic necrosis virus (IHNV) are listed as the first and second class infectious diseases of imported animals in the People's Republic of China. VHS and IHN are the second-class and third-class infectious diseases stipulated in the "Animal Epidemic Prevention Law of the People's Republic of China" respectively. These three diseases are listed in the OIE list of the International Office of Epizootics. Both IHN and VHS viruses belong to the single-stran...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04C12Q1/68
Inventor 段宏安张睿姚燕林王海涛周毅李金华刘小琴
Owner LIANYUN PORT IMMIGRATION INSPECTION & QUARANTINE BUREAU PEOPLES REPUBLIC OF CHINA