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Method for detecting Nuoda Virus mucleic-acid point cross-breeding of luoshi shrimp in natural pond

A technology of Nodavirus and Macrobrachium rosenbergii is applied in the diagnosis of the pathogen of Macrobrachium rosenbergii muscle white turbidity, and the nucleic acid dot hybridization detection of Macrobrachium rosenbergii, which can solve the problems of low sensitivity and achieve high sensitivity and sample processing. Simple procedure and low cost effect

Inactive Publication Date: 2003-08-27
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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Problems solved by technology

Data (4) reported an immunological detection method for Macrobrachium rosenbergii Nodavirus, which is simple and fast, but has low sensitivity

Method used

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Embodiment Construction

[0019] Specific steps are as follows:

[0020] (1) Preparation kit: The kit contains all the necessary components in the detection procedure, including viral nucleic acid probes, sample processing fluid, hybridization fluid, blocking fluid, anti-digoxigenin antibody, washing fluid, color reagent, positive control, Negative control, 1.5ml centrifuge tube, hybridization membrane, hybridization belt, grinding rod.

[0021] (2) Preparation of probes: The preparation of digoxin probes was carried out by conventional PCR methods, and the PCR products were labeled with digoxin. The amplification primers were derived from the plasmid universal primers to construct the Macrobrachium rosenbergii Noda virus cDNA library.

[0022] (3) Sample treatment: Weigh the sample (the whole seedling or the gill of the adult shrimp), put it into a 1.5ml centrifuge tube, add the sample treatment solution (1:10 weight / volume), the sample treatment solution is treated with diethyl pyrocarbonate Buffer solut...

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Abstract

The invention discloses a method for checking nucleic acid point crossbreed of rose zhaoxianuoda virus, the steps are; at first, the producing of probe; the second is the treatment of samples; the third is preliminary crossing and crossing; the fourth is rinsing; the fifth is obturation; then sixth is antibody antigen reaction; the seventh is rinsing; the eighth is color-reaction; finally ends up the reaction.

Description

Technical field: [0001] The invention belongs to the technical field of shrimp disease detection, and specifically relates to a method for detecting the nucleic acid dot hybridization of Macrobrachium rosenbergii Noda virus, which is suitable for the diagnosis of the pathogen of Macrobrachium rosenbergii muscle white turbidity. Background technique: [0002] Macrobrachium rosenbergii muscle whitish disease has been the main disease affecting the farming industry of Macrobrachium rosenbergii in Southeast Asia and the Caribbean since the 1990s (Nash et al., 1987; Tung et al., 1999; Arcier et al., 1999) . The target of the disease is the seedlings in the nursery, and the muscles of the diseased seedlings are cloudy. The disease caused a large number of deaths of shrimp larvae in a short period of time. According to the "Monthly Report on Aquaculture Animal and Plant Diseases" edited by the National Aquatic Technology Extension Station of the Fisheries Bureau of the Ministry of Agric...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/34C12Q1/68
Inventor 石正丽宝纳米・简・罗伯特
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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