Quantitative detection method for halomonas strains by adopting indirect enzyme-linked immunosordent assay

A quantitative detection method, Halomonas technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of difficult sample detection, time-consuming, laborious, and randomness, etc., and achieve easy operation, simple operation process, and specificity strong effect

Active Publication Date: 2015-05-27
OCEAN UNIV OF CHINA
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  • Application Information

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Problems solved by technology

At present, the detection of Halomonas is mainly through traditional detection techniques, such as plate counting method and direct microscope counting me

Method used

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  • Quantitative detection method for halomonas strains by adopting indirect enzyme-linked immunosordent assay
  • Quantitative detection method for halomonas strains by adopting indirect enzyme-linked immunosordent assay
  • Quantitative detection method for halomonas strains by adopting indirect enzyme-linked immunosordent assay

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Experimental program
Comparison scheme
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Embodiment Construction

[0042] Further illustrate the present invention below by embodiment

[0043] 1. Experimental animals:

[0044] 5 healthy male rabbits (more than 6 months old, weighing 2-3kg, fed pelleted mixed feed, temperature 16-28°C, temporarily raised for one week)

[0045] 2. Main instruments:

[0046] RO15 pure water machine, Shanghai Lishen Scientific Instrument Co., Ltd.

[0047] Neofuge 15R desktop centrifuge, Shanghai Lishen Scientific Instrument Co., Ltd.

[0048] TS-1 shaker, Jiangsu Haimen Qilin Medical Instrument Factory

[0049] SPECTRA MAX 190 Microplate Reader, Molecular Devices

[0050]Adjustable pipette, Effendorf

[0051] 3. Reagents

[0052] Horseradish peroxidase-labeled goat anti-rabbit IgG-HRP, Beijing Hanpu Medical and Biological Research Institute

[0053] Tetraphenylbenzidine (TMB) Chromogenic Solution, Beijing Hanpu Medical and Biological Research Institute

[0054] Other reagents are analytical reagents

[0055] 4. Steps:

[0056] 1. Preparation of Halom...

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Abstract

The invention provides a quantitative detection method for halomonas strains by adopting an indirect enzyme-linked immunosordent assay. The quantitative detection method comprises the following steps: (1) preparing an O antigen of halomonas; (2) preparing a polyclonal antibody of the halomonas, namely with the O antigen of the halomonas as an immunizing antigen, immunizing a New Zealand rabbit, and preparing the polyclonal antibody of the halomonas by adopting an existing polyclonal antibody preparation and purification technology; and (3) carrying out an indirect ELISA operation method, namely coating the antigen with an elisa plate, and staying overnight at 4 DEG C, washing the plate, adding confining liquid, washing the plate, and adding the antibody, washing the plate, and adding a second antibody, washing the plate, and carrying out chromogenic reaction, and measuring the light absorption value, namely OD450 at a wavelength of 450nm by using a microplate reader. The quantitative detection method for the halomonas strains by adopting the indirect enzyme-linked immunosordent assay is simple and convenient in operation flow and is easy to operate, the detection sensitivity can reach 105cfu/mL, and antiserum generates no cross reaction with other bacteria, so that the quantitative detection method is a fast and efficient detection method with high sensitivity and strong specificity, promotes the application of probiotics on aquatic products and has the certain practical application significance.

Description

technical field [0001] The invention relates to a method for detecting the content of Halomonas, more specifically an indirect ELISA quantitative detection method for Halomonas strains, belonging to the technical field of immune detection. Background technique [0002] In the process of applying probiotics to improve water quality, in order to judge the improvement of water quality by different concentrations of beneficial bacteria, and to understand the changes in the number of probiotics in the water, it is often necessary to quantitatively detect them. Therefore, it is necessary to establish accurate quantitative detection technology. [0003] Halomonas has a wide range of adaptability to temperature, salinity and oxygen, and has strong adaptability. It has important application value in aquaculture water purification, sewage treatment and bioremediation. At present, the detection of Halomonas is mainly through traditional detection techniques, such as plate counting met...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/535
CPCG01N33/535G01N33/56911
Inventor 李贇苏真真潘鲁青
Owner OCEAN UNIV OF CHINA
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