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Fluorescence quantitative PCR detection kit and detection method for prawn infectious myonecrosis viruses

A detection kit and technology for muscle necrosis, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganism measurement/testing, etc., can solve the problem of low sensitivity, achieve method sensitivity, high promotion and application value, and easy operation Effect

Inactive Publication Date: 2012-07-11
LUDONG UNIVERSITY
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Problems solved by technology

[0004] The purpose of the present invention is to solve the problem that the sensitivity of existing fluorescent quantitative PCR detection method is not high, provide the fluorescent quantitative PCR detection kit of prawn infectious myonecrosis virus, also provide the detection method that utilizes this kit to detect simultaneously, The detection method is sensitive, specific and easy to operate

Method used

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  • Fluorescence quantitative PCR detection kit and detection method for prawn infectious myonecrosis viruses
  • Fluorescence quantitative PCR detection kit and detection method for prawn infectious myonecrosis viruses
  • Fluorescence quantitative PCR detection kit and detection method for prawn infectious myonecrosis viruses

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Embodiment 1

[0027] The fluorescent quantitative PCR detection kit for shrimp infectious myonecrosis virus comprises a box body 1, and a tube hole support 5 is arranged in the box body 1, and the following pipe fittings are arranged on the tube hole support 5:

[0028] (1) Fluorescent quantitative PCR reaction solution tube 6, which contains 2× fluorescent quantitative PCR buffer, a mixture of positive and negative primers for shrimp infectious myonecrosis virus and fluorescently labeled TaqMan probes;

[0029] Forward primer sequence: 5'- GATGG CAGCA CGTCA AAATG -3';

[0030] Reverse primer sequence: 5'- CAGCT GTTCC TGGGA CTTCC T -3';

[0031] TaqMan probe sequence: 5'-CTATT TCCTC CACTA ATTCA GAGC-3', the 5' end of the probe sequence is labeled with FAM, and the 3' end is labeled with TAMRA;

[0032] (2), deionized water pipe 3, filled with sterile deionized water;

[0033] (3) Negative control tube 4, containing SPF (specific pathogen free) prawn nucleic acid;

[0034] (4), standard q...

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Abstract

The invention relates to a fluorescence quantitative PCR detection kit and a detection method for a prawn infectious myonecrosis viruses, which belongs to the technical field of detection of pathogeny of marine organisms. The fluorescence quantitative PCR detection kit comprises a box, a pore support is provided in the box, and the following pipe fittings are placed on the pore support: (1) a fluorescence quantitative PCR reaction solution pipe which is filled with a forward primer sequence, a reverse primer sequence and a TaqMan probe sequence; (2) a deionized water pipe which is filled with aseptic deionized water; (3) a negative control pipe which is filled with SPF (specific pathogen free) prawn nucleic acid; and (4) standard substance pipes which are respectively filled with infectious myonecrosis virus 10<7>, 10<6>, 10<5>, 10<4>, 10<3>, 10<2> and 10 and one copied cDNA positive clone. The invention has high popularization and application values, and the detection method is sensitive and specific and is convenient to operate.

Description

Technical field [0001] The invention involves detection kits and detection methods for shrimp infectious muscle necrosis virus, which belongs to the field of marine biological pathogenic testing technology. Background technique [0002] Infectious muscle necrosis is a new viral infectious disease that is harmful to shrimp production in Brazil in 2002. In 2004, Dr. Lightner D V, the University of Arizona, found that the disease was caused by a new virus and will be caused by a virusNamed in Infectious Myonecrosis Virus (IMNV).This new type of virus is a double -chain RNA virus, which belongs to the overall virus.The susceptible pairing of shrimp infectious muscle necrotic virus in shrimp is South whitening shrimp, which is mainly infected with young shrimp infected with 60-80 days.The virus can cause necrosis of the infected disease to the whole body muscle tissue of shrimp. In general, the pathological death is slow and the mortality rate is not high. However, the entire breeding...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34C12Q1/70C12Q1/68C12R1/93
Inventor 闫冬春刘鸿玲
Owner LUDONG UNIVERSITY
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