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Method for hybrid detecting nucleic acid complex point of shrimp viral

A virus nucleic acid and compound point technology, which is applied in biochemical equipment and methods, measuring devices, and microbial determination/inspection, etc., can solve problems such as complex operation procedures, and achieve the effects of simple sample processing, high sensitivity and low cost.

Inactive Publication Date: 2003-10-29
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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AI Technical Summary

Problems solved by technology

In situ hybridization has high sensitivity, but the operation procedure is complicated, which is suitable for the intracellular localization of viruses

Method used

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Examples

Experimental program
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Embodiment Construction

[0024] Specific steps are as follows:

[0025] (1) Prepare the kit: the kit contains the necessary components in all detection procedures, including viral nucleic acid probe, sample processing solution, hybridization solution, blocking solution, anti-digoxigenin antibody, anti-biotin antibody, washing solution, color development Reagent, positive control, negative control, 1.5ml centrifuge tube, hybridization membrane, hybridization bag, grinding stick.

[0026] (2) Preparation of probes: Digoxigenin probes and biotin probes were prepared using conventional PCR methods, and PCR products were labeled with digoxin and biotin, respectively. The amplification primers were designed from the sequences of the shrimp white spot syndrome probe and the Torah syndrome virus probe, and the amplification template was from the viral nucleic acid.

[0027] (3) Sample treatment: Weigh the sample (the whole seedling or the gills of adult shrimp), put it into a 1.5ml centrifuge tube, add the s...

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Abstract

A method for detecting the virus of prawn by nucleic acid hybridization includes preparing probes, treating specimen, pre-hybridizing, hybridizing, washing, closing, antibody-antigen reaction, washing, two-step color development reaction, and terminating reaction. It can detect two kinds of virus at same time. Its advantages are high sensitivity and low cost.

Description

technical field [0001] The invention belongs to the technical field of shrimp disease detection, and in particular relates to a method for detecting prawn virus nucleic acid complex point hybridization, which is suitable for the diagnosis of the pathogens of prawn white spot syndrome and Torah syndrome. Background technique [0002] White Spot Syndrome Virus (WSSV) of prawn (Penaeus) was first discovered in Taiwan, China in the 1990s. It is one of the main virus pathogens that cause serious economic losses in prawn farming. It can infect most prawn species, and the fatality rate is Up to 90-100%. The main symptom is that the body color of prawns is red, and white spots appear under the skin, carapace and appendages. It can also infect other freshwater and marine crustaceans. Taura syndrome virus (Taura Syndrome Virus, TSV) was first discovered in Penaeus vannamei in 1992, and the cumulative mortality caused by infection of cultured Penaeus van...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/34C12Q1/68G01N33/569
Inventor 石正丽袁军法
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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