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Colloidal gold visual detection method for Micropterus salmoides iridovirus and probe for method

A technology of iridescent virus and detection method, which is applied in the field of colloidal gold visual detection of iridescent virus of the largemouth bass frog virus genus, can solve the problems that no research involves the application of AuNPs, and achieve the effect of being suitable for on-site detection, low cost and simple method

Pending Publication Date: 2020-01-17
ZHEJIANG GONGSHANG UNIVERSITY
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  • Abstract
  • Description
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Problems solved by technology

[0006] However, there are currently no studies involving the application of AuNPs in LMBV detection

Method used

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  • Colloidal gold visual detection method for Micropterus salmoides iridovirus and probe for method
  • Colloidal gold visual detection method for Micropterus salmoides iridovirus and probe for method
  • Colloidal gold visual detection method for Micropterus salmoides iridovirus and probe for method

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Embodiment 1

[0051] Embodiment 1, a kind of colloidal gold visual detection method of largemouth bass frog virus genus iridescent virus, carries out the following steps successively:

[0052] ①Design and synthesis of largemouth bass iridescent virus nucleic acid probe:

[0053]

[0054] ② Preparation of colloidal gold:

[0055] Heat 100mL of auric acid solution (0.01%, ie, 0.01g / 100ml) to boiling, add 3.5mL of 1% (1g / 100ml) sodium citrate solution under stirring conditions, the system changes from yellow to wine red, continue heating and boiling After 15 minutes, remove the heat source and stir until cooled to room temperature (stirring time is about 20-30 minutes) to obtain colloidal gold (AuNPs solution), which can be stored at 4°C.

[0056] ③Incubation of nucleic acid probe-colloidal gold complex:

[0057] Mix 20 μL (10 μM) of the probe with 1 mL of colloidal gold and incubate at 37°C for 24 hours. During this period, invert and mix several times. After the incubation, the AuNPs-pr...

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Abstract

The invention discloses a colloidal gold visual detection method for Micropterus salmoides Ranavirus iridovirus and a probe for the method; the method comprises: incubating the probe with colloidal gold to obtain AuNPs-probe; extracting virus DNA in Micropterus salmoides to be detected; mixing the virus DNA with AuNPs-probe, culturing at 37 DEG C, and adding NaCl solution; observing the color change of the mixed solution 2 minutes later; if the color of the mixed solution is still red, determining that the result is negative, and if the color of the mixed solution is gray blue, determining that the result of the sample is positive. The method is simple, low in cost, high in speed and reliable, and is more suitable for on-site detection. The invention provides the effective method for rapiddetection of Micropterus salmoides iridovirus and provides technical guarantee for aquaculture.

Description

technical field [0001] The invention relates to a colloidal gold visual detection method for largemouth bass frog virus genus iridescent virus based on a nucleic acid probe. Background technique [0002] The scientific name of the California perch is the largemouth bass (Micopterus salmoides), which is native to the Mississippi River Basin in North America. Micropterus). California perch has strong adaptability, and has the advantages of large body, fast growth, easy catch, and short breeding cycle. In addition, the meat is delicious and tender, without intermuscular spines, and has a beautiful appearance. It is welcomed by farmers and consumers all over the world. Domestic Guangdong Province introduced California perch for the first time in 1983, and has now become an important and precious species of freshwater aquaculture in my country. It is widely cultivated in provinces south of the Yangtze River (such as Guangdong, Zhejiang, Jiangsu and Sichuan, etc.), with a total a...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6816C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6816C12Q2563/137C12Q2563/155
Inventor 冯俊丽朱勤超金仁耀
Owner ZHEJIANG GONGSHANG UNIVERSITY
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