Method for quantitatively detecting white spot syndrome virus through TaqMan probe

A technique for quantitative detection of vitiligo syndrome, applied in the field of molecular biology, can solve the problems of time-consuming results, discrepancies, etc., and achieve the effects of short time, high detection sensitivity, and good application prospects.

Inactive Publication Date: 2018-07-31
SHANGHAI OCEAN UNIV
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  • Description
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Problems solved by technology

A single detection method usually has defects, and simultaneous detection by multiple methods is not only time-consuming and the results are different, so it is necessary to establish an accurate and quantitative method for the detection of white spot syndrome virus in shrimp

Method used

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  • Method for quantitatively detecting white spot syndrome virus through TaqMan probe
  • Method for quantitatively detecting white spot syndrome virus through TaqMan probe

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Embodiment 1

[0021] A method for quantitatively detecting prawn white spot syndrome virus using TaqMan probes, comprising the steps of:

[0022] (1) Collect 50 fresh muscle samples of shrimp larvae, put them into alcohol immediately after collection and store them at 4°C.

[0023] Viral nucleic acid extraction: The DNA extraction kit of Tiangen Company was used, and the steps were carried out by referring to the instructions in the kit.

[0024] (2) Design primers and TaqMan probes

[0025] The design of the primer probe was searched for the related sequence of White Spot Syndrome Virus (WSSV) on NCBI, compared with BioEdit software, analyzed the conserved region and specific region of the sequence, determined the primer and probe, and then used Primer Premier5 for primer detection. Needle assessment, initial screening. After synthesizing primers and probes, use PCR to verify whether the target fragment is amplified and the specificity of amplification, and further determine feasible pri...

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Abstract

The invention relates to a method for quantitatively detecting white spot syndrome virus through a TaqMan probe. The method comprises the steps: firstly, extracting genome DNA of a sample to be detected; then designing a primer and the TaqMan probe; finally, performing fluorogenic quantitative PCR detection on the genome DNA of the sample to be detected through the designed primer and the designedTaqMan probe. The detection method disclosed by the invention has the advantages of high detection flexibility and good specificity, short used time, no false positive situation generation, application to molecular disease detection and early warning of prawn farming in China and good application prospect; furthermore, the lowest copy number which can be detected by the detection method is 7.8.

Description

technical field [0001] The invention relates to a method for quantitatively detecting prawn white spot syndrome virus (WSSV), which belongs to the technical field of molecular biology. Background technique [0002] Shrimp white spot syndrome virus (WSSV) has become the most dangerous source of infection in shrimp farming since it was first discovered in 1992, and it has a wide host range in decapod crustaceans. The WSSV genome is a uniformly distributed circular dsDNA molecule with different genome sizes in different strains, and it is one of the largest viral genomes sequenced so far. The full genome sequences of three WSSV strains have been determined, including: Thailand strain 293kbp, Chinese strain 305kbp and Taiwan strain 307kbp, the similarity between them is 99.3%. Different segments of the WSSV genome have important sequence differences, which can be used to determine the origin and spread of WSSV strains, as well as to distinguish strains in the wild, and may also...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6851
CPCC12Q1/6851C12Q1/701C12Q2531/113C12Q2561/101
Inventor 刘红李云于晓丹戴习林朱其建周微微
Owner SHANGHAI OCEAN UNIV
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