Luminescence immune detection method based on recombinant antigen carrying His tag

Abstract

The invention provides a luminescence immune detection method based on a recombinant antigen carrying a His tag. His-tag-carried recombinant protein serving as a known antigen, His antibody coating resistant luminescent microspheres, biotin labeled second antibodies and streptavidin labeled light sensing microspheres jointly form a detection reagent. According to the method, the recombinant protein carrying the His tag is taken as the known antigen, the method has the most important characteristic that the known antigen is arranged in liquid-phase three-dimensional space, which is essentially different from an immobilized antigen in the prior art. Due to the change, the technical problems caused by the immobilization process in a traditional antibody detection method are effectively solved. Besides, a homogeneous-phase luminescence immunoassay system based on active oxygen energy transmission is adopted and has higher analysis sensitivity and precision.

Description

technical field [0001] The invention belongs to the technical field of antibody detection in the field of medical testing, mainly including allergen antibodies, pathogen antibodies and autoantibodies, and in particular relates to a luminescent immunodetection method based on recombinant antigens carrying His tags. Background technique [0002] Immunochemistry (antigen-antibody reaction) technology is an important method for detecting specific antibodies, that is, using known antigens to detect unknown antibodies. Enzyme-linked immunosorbent assay (ELISA) indirect method is often used to detect the above antibodies, and enzyme-linked dot blot assay (dot-ELISA) can also be used to simultaneously coat multiple related antigens in a "linear manner" to achieve simultaneous detection of multiple antibodies ( Such as antinuclear antibody profile, HIV infection confirmation test, etc.). The basic principles of the above two methods are as follows: a solid phase carrier (polystyrene...

AI Technical Summary

Problems solved by technology

[0006] Third, the preparation process and use process are complex. Regardless of ELISA or ELISA dot blot test, the process of immobilization of antigens is a key link in the production of diagnostic reagents. The process of antigen coating-sealing-drying is more complicated, and standardization needs Standard operating procedures and strictly enforced

From the user's point of view, the use of the above two detection reagents is also relatively complicated. Clinical specimens need to be diluted, two incubations and multiple washings will take a long time, and the labeled antibody (enzyme) is affected by various factors, etc. User-friendly

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