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Method for detecting Streptococcus agalactiae from culture water body to prevent Tilapia mossambica from suffering from diseases

A technology for streptococcus lactis and aquaculture water, which is applied in the field of DNA fragment detection, can solve the problems of high cost, antibiotic residues in fish, and LAMP prone to false positives, and achieve the effect of prevention

Pending Publication Date: 2021-02-05
GUANGXI UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these technologies also have certain deficiencies, such as 16S sequencing technology is difficult to distinguish species with high homology; fluorescent antibody technology and ELISA are expensive and have complicated operation steps; LAMP is prone to false positives; quantitative PCR requires expensive instruments , professional operation and high cost; there will be interference between two or more pairs of primers in double and multiplex PCR, etc.
[0010] The existing prevention and control technology of Streptococcus tilapia is to wait until the fish body is on the disease before starting the treatment. At this stage of the disease, the pathogen has already existed in large quantities in the water body and fish body, and it is very difficult to cure it. Generally, long-term medication is required to control it. It will cause problems such as antibiotic residues in fish and drug residues in water

Method used

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  • Method for detecting Streptococcus agalactiae from culture water body to prevent Tilapia mossambica from suffering from diseases
  • Method for detecting Streptococcus agalactiae from culture water body to prevent Tilapia mossambica from suffering from diseases
  • Method for detecting Streptococcus agalactiae from culture water body to prevent Tilapia mossambica from suffering from diseases

Examples

Experimental program
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Effect test

Embodiment 1

[0038] The method for detecting streptococcus agalactiae to prevent disease of tilapia from culture water body comprises the steps:

[0039] 1) Take 1000ml of pond water, put it into four 250ml centrifuge tubes on average, and centrifuge at 8000rpm for 10min;

[0040] 2) Remove the supernatant, leave 10ml of liquid in each tube, resuspend evenly with the precipitate, put 4 tubes of resuspension into a 50ml centrifuge tube, centrifuge at 8000rpm for 10min, remove the supernatant, leave 1-2ml of liquid, and mix with the precipitate Resuspend evenly;

[0041] 3) Extract the total DNA of the resuspension, and use the extracted total DNA as a template to perform PCR amplification to amplify the cfb gene fragment of Streptococcus agalactiae; the primer sequences used are P1: 5-TATCTGGAACTCTAGTGGCTG-3, P2: 5- TGACGTTAAGTACTTTTTTATC-3; the reaction conditions were pre-denaturation at 94°C for 5 minutes; denaturation at 94°C for 30s, annealing at 52°C for 30s, extension at 72°C for 60...

Embodiment 2

[0048] Methods for detecting Streptococcus agalactiae from aquaculture water to prevent disease in tilapia:

[0049] The difference with embodiment 1 is that step 5) uses chlorine dioxide to carry out disinfection, and consumption is: 1 mu of water body (according to 1 meter water depth) uses the chlorine dioxide effervescent tablet 400g of effective content 7%;

[0050] Others are the same as embodiment 1.

Embodiment 3

[0052] Methods for detecting Streptococcus agalactiae from aquaculture water to prevent disease in tilapia:

[0053] The difference with embodiment 1 is that step 5) uses chlorine dioxide to carry out disinfection, and consumption is: 1 mu of water body (according to 1 meter water depth) uses the chlorine dioxide effervescent tablet 400g of effective content 8%;

[0054] Others are the same as embodiment 1.

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Abstract

The invention discloses a method for detecting Streptococcus agalactiae from a culture water body to prevent Tilapia mossambica from suffering from diseases. The method is a method for detecting Streptococcus agalactiae from a water body with ultrahigh sensitivity, and the sensitivity of the method can reach 1-10 CFU / 10 ml. By means of the method, whether Streptococcus agalactiae exists or not canbe detected from the water body before Streptococcus agalactiae infects the fish body on a large scale, if yes, measures such as pond disinfection and antibiotic feeding are taken immediately to prevent diseases, and the treatment means are implemented at the earliest stage of disease development, so that the effect of prevent Tilapia mossambica from suffering from diseases is very obvious.

Description

【Technical field】 [0001] The invention relates to the technical field of DNA fragment detection, in particular to a method for detecting streptococcus agalactiae from aquaculture water to prevent tilapia from getting sick. 【Background technique】 [0002] Tilapia is one of the most widely farmed fish in the world. It has a series of advantages such as fast growth, strong disease resistance and stress resistance, good meat quality, strong fecundity, wide adaptability and high population yield. The global production is only in In 2013 it reached 4.5 million tons. In my country, tilapia has become the largest export-oriented farmed fish in the southern provinces, and its output accounts for 50% of the world's total. The tilapia industry is of great significance for increasing employment opportunities and increasing farmers' income. [0003] Although the tolerance of tilapia in poor water quality is better than that of other freshwater fish, with the extensive development of aqu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/04C12Q1/686C12N15/11C12R1/46
CPCC12Q1/689C12Q1/686C12Q2561/113C12Q2563/107
Inventor 易弋罗福广佀再勇黄杰龙秀锋文衍红
Owner GUANGXI UNIVERSITY OF TECHNOLOGY