A kind of indirect ELISA quantitative detection method of halomonas strain

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...

AI Technical Summary

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 method. The disadvantages are large randomness, long time-consuming, time-consuming and laborious, and it is difficult to use for a large number of samples.

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