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Method for quantitatively detecting adhesion degree of bacterial genus on surfaces of coating layers in early contamination stage by utilizing genus-specific primers

A quantitative detection and specific technology, applied in the biological field, can solve the problems of insufficient research on the early attachment process of fouling organisms

Inactive Publication Date: 2018-11-13
HARBIN INST OF TECH AT WEIHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, most of the researches on fouling organisms focus on the ecology of fouling organisms and antifouling technology, and the research on the early attachment process of fouling organisms is still insufficient.

Method used

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  • Method for quantitatively detecting adhesion degree of bacterial genus on surfaces of coating layers in early contamination stage by utilizing genus-specific primers
  • Method for quantitatively detecting adhesion degree of bacterial genus on surfaces of coating layers in early contamination stage by utilizing genus-specific primers
  • Method for quantitatively detecting adhesion degree of bacterial genus on surfaces of coating layers in early contamination stage by utilizing genus-specific primers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment example 1

[0047] Implementation Case 1. Preparation of Marine Fouling Samples

[0048] In order to explore the microbial community on the surface of the antifouling coating added with different nanomaterials, the hanging plate test was carried out to simulate the state of the hull in seawater, so as to obtain marine fouling biological community samples.

[0049] (1) Selection of nanomaterials: the selected nanomaterials are as shown in Table 5:

[0050] Table 5 Nanomaterials

[0051]

[0052] (2) Coating: Mix the 4 nanomaterials shown in the above table with different types and different concentrations of primers and mix them evenly on the surface of the steel plate. There are 11 types of surfaces in total, numbered a-k, and dry at room temperature. The material and thickness of the steel plates used in the experiment are consistent with those of the ship bottom manufactured by Weihai Shipyard, which can basically achieve the effect of simulating the bottom of the ship in seawater. ...

Embodiment example 2

[0056] Example 2. Using specific primer amplification results to semi-quantitatively assess the attachment of 6 genera of microorganisms in seawater fouling samples

[0057] The screened 6 pairs of genus-specific primers were applied to the microbial communities on the surface of oyster growth-promoting and oyster-inhibiting materials, and the differences in community biodiversity were determined based on PCR amplification results and corresponding primer information. For this reason, an equal volume of eluent was taken out from the tubes storing the eluent of each small steel plate sample into a new tube, and the samples were labeled. The first sampling samples were labeled a1, b1, d2, e2, f2, g2, h2, i2, j2, k2, empty, the second sample number is a2, b2, d2, f2, g2, empty, the third sample number is a3, b3, c3, d3, e3, f3, g3, h3, i3, j3, k3 ("empty" represents the blank group, that is, a small steel plate sample without nanomaterials on the surface); the metagenomic group o...

Embodiment example 3

[0064] Implementation case 3. Several analyzes of the characteristics of microbial communities on the surface of steel plates

[0065] The 6 pairs of specific primers screened by the current design and the PCR amplification results of all samples on the surface of oyster growth-promoting nanomaterials and inhibiting nanomaterials, including whether amplified bands are amplified and the brightness of the bands, are all integrated for community analysis . According to the analysis results, there were Flavobacterium, Glaciecola, Jannaschia, Rhodococcus, sub Growth of Sulfitobacter. According to the band brightness of the amplified product, it can be clearly seen that Jannaschia (Jannaschia) grows more vigorously and has more biomass on the surface of the material that promotes the growth of oysters than on the surface of the material that inhibits the growth of oysters. There was no significant difference in the growth and reproduction degrees of the other four genera on the su...

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Abstract

The invention belongs to the technical field of biology, and relates to a method for quantitatively detecting the adhesion degree of six bacterial genuses on the surfaces of different coating layers in an early contamination stage by utilizing genus-specific primers. The method comprises the following steps: (1) designing corresponding specific primers by taking six bacterial genuses (Acinetobacter, Flavobacterium, Glaciecola, Jannaschia, Rhodococcus and Sulfitobacter) detected in samples on the surface of a marine hanging plate as objects; (2) collecting seawater contaminated samples on the surfaces of different coating layers in the early contamination stage, and carrying out metagenome extraction; (3) carrying out PCR (Polymerase Chain Amplification) for detecting the adhesion degree ofthe six bacterial genuses by utilizing the specific primers. Species and genus-specific primers are designed based on the specific primers in whole genome sequences of ocean contaminated prokaryotes,and an efficient and fast technological means is provided for detecting the dynamic change of a contaminated microbial community structure in the samples at the level of species and genus.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for quantitatively detecting the adhesion degree of six kinds of fouling microbial genera on different coating surfaces in the early stage of fouling by using genus-specific primers. This method provides an efficient and rapid technical means for detecting the dynamic changes of the fouling microbial community structure in samples, and can be applied to detect the adhesion degree of fouling microorganisms on different marine engineering surface coatings. Background technique [0002] Marine biofouling has brought serious harm to the development of the marine industry around the world, greatly restricting the development and utilization of marine resources by humans, and marine antifouling has become an important issue that needs to be solved urgently. Many studies have shown that the marine early-fouling microbial community has an important impact on the qualitative attachment...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/06
CPCC12Q1/689
Inventor 谷庆泽张艾涵张治洲宋媛李凯强
Owner HARBIN INST OF TECH AT WEIHAI