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Visible rapid detection kit and detection method for prawn white spot syndrome virus

A technology for vitiligo syndrome and detection kits, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as false positives, false negatives, and sufficient nucleic acid extraction

Inactive Publication Date: 2016-03-09
天津市水生动物疫病预防控制中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, this technology has been applied to the detection of WSSV, but due to the rapid nucleic acid extraction method may not be able to fully extract the nucleic acid, resulting in false negatives, and the use of dyes such as SYB-Green may produce false positives, etc., hindering LAMP Popularization and Application of Detection Technology in the Process of Shrimp Breeding

Method used

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  • Visible rapid detection kit and detection method for prawn white spot syndrome virus
  • Visible rapid detection kit and detection method for prawn white spot syndrome virus
  • Visible rapid detection kit and detection method for prawn white spot syndrome virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] A visual rapid detection kit for prawn white spot syndrome virus, comprising:

[0045]Reagent A, prepared by the following method: add cetyltrimethylammonium bromide, NaCl, ethylenediaminetetraacetic acid and hydroxymethylaminomethane-hydrochloric acid to 100ml to obtain a solution, cetyltrimethylammonium bromide in the solution The grams of methylammonium bromide is 1.5g, the concentration of NaCl is 1.3M, the concentration of ethylenediaminetetraacetic acid is 15mM, and the concentration of hydroxymethylaminomethane-hydrochloric acid is 15mM; adjust pH=7.5 to obtain reagent A;

[0046] Reagent B: proteinase K aqueous solution with a concentration of 20mg / ml;

[0047] Reagent C: 100mg nucleic acid extraction magnetic beads resuspended in 1ml double distilled water;

[0048] Reagent D: isopropanol;

[0049] Reagent E: 65% ethanol water solution by volume;

[0050] Reagent F: sterile double distilled water;

[0051] Reagent G: prawn white spot syndrome virus positive...

Embodiment 2

[0055] A visual rapid detection kit for prawn white spot syndrome virus, comprising:

[0056] Reagent A, prepared by the following method: add cetyltrimethylammonium bromide, NaCl, ethylenediaminetetraacetic acid and hydroxymethylaminomethane-hydrochloric acid to 100ml to obtain a solution, cetyltrimethylammonium bromide in the solution The grams of methylammonium bromide is 2.5g, the concentration of NaCl is 1.4M, the concentration of ethylenediaminetetraacetic acid is 20mM, and the concentration of hydroxymethylaminomethane-hydrochloric acid is 20mM; adjust pH=7.5 to obtain reagent A;

[0057] Reagent B: proteinase K aqueous solution with a concentration of 25 mg / ml;

[0058] Reagent C, reagent D are the same as reagent C and reagent D of embodiment 1;

[0059] Reagent E: ethanol aqueous solution with a volume percentage concentration of 70%;

[0060] Reagent F, reagent G, the detection tube that reagent H is housed are the same as the reagent F, reagent G, and the detecti...

Embodiment 3

[0063] A visual rapid detection kit for prawn white spot syndrome virus, comprising:

[0064] Reagent A, prepared by the following method: add cetyltrimethylammonium bromide, NaCl, ethylenediaminetetraacetic acid and hydroxymethylaminomethane-hydrochloric acid to 100ml to obtain a solution, cetyltrimethylammonium bromide in the solution The number of grams of methyl ammonium bromide is 3g, the concentration of NaCl is 1.5M, the concentration of ethylenediaminetetraacetic acid is 25mM, and the concentration of hydroxymethylaminomethane-hydrochloric acid is 25mM; adjust pH=7.5 to obtain reagent A;

[0065] Reagent B: proteinase K aqueous solution with a concentration of 30mg / ml;

[0066] Reagent C, reagent D are the same as reagent C and reagent D of embodiment 1;

[0067] Reagent E: a 75% ethanol aqueous solution with a volume percent concentration;

[0068] Reagent F, reagent G, the detection tube that reagent H is housed are the same as the reagent F, reagent G, and the det...

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Abstract

The invention discloses a visual rapid detection kit and a detection method for white spot syndrome viruses. The detection kit comprises a reagent A, a reagent B, a reagent C, a reagent D, a reagent E, a reagent F, a reagent G and a detection tube filled with a reagent H, wherein the reagent A is a solution prepared by adding cetyl trimethyl ammonium bromide, NaC1, ethylenediamine tetraacetic acid and hydroxy methyl amino methane-hydrochloric acid in water to 100ml; the reagent B is a proteinase K aqueous solution; the reagent C is a solution prepared by re-suspending magnetic beads for nucleic acid extraction in double distilled water; the reagent D is isopropanol; the reagent E is an ethanol water solution; the reagent F is sterilized double distilled water; the reagent G is positive plasmids of white spot syndrome viruses. The whole operation process of the kit is very simple and convenient, WSSV (white spot syndrome viruses) detection can be completed within 1.5-1.2 hours, any high-end equipment is not needed, and only one water bath kettle or metal bath is needed. The kit is suitable for being popularized and applied in the process of practical production.

Description

technical field [0001] The invention relates to the detection technology of pathogens in aquaculture, in particular to a visual and rapid detection kit and detection method for white spot syndrome virus (WhiteSpot Syndrome Virus, WSSV) of prawns. Background technique [0002] Shrimp white spot syndrome virus is a double-stranded DNA virus with an envelope, which can cause 100% mortality in prawns within 5-7 days, causing serious economic losses to the prawn farming industry. The host range of white spot virus infection is very wide, not only infecting various wild and farmed prawns, but also infecting other aquatic crustaceans such as crabs, crayfish and lobsters, etc., especially the most serious impact on prawn populations. In 2009, prawn white spot syndrome was listed as a first-class disease of aquatic organisms by the Ministry of Agriculture's "List of Types I, II, and III Animal Diseases". There is currently no effective treatment for prawn white spot syndrome. Early ...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/6844C12Q1/70C12Q2531/119C12Q2563/107
Inventor 孙金生薛淑霞孙妍耿绪云
Owner 天津市水生动物疫病预防控制中心