A human anti-AQP4 autoantibody detection material and preparation method thereof

Abstract

The invention provides a human body anti-AQP4 autoantibodies detection material and a preparation method thereof. The preparation method comprises the steps of firstly acquiring M1 and M23 subtype AQP4 overall length target genes, dyeing into an HEK293 cell, and building an HEK293 / pCI-neo-M1-AQP4 cell and an HEK293 / pCI-neo-M23-AQP4 cell expressing M1 and M23 subtype AQP4; extracting an AQP4-cell membrane complex of the cells; finally curing the AQP4-cell membrane complex on a carrier, and obtaining the human body anti-AQP4 autoantibodies detection material. According to the human body anti-AQP4 autoantibodies detection material and the preparation method thereof provided by the invention, the extracted AQP4-cell membrane complex is used as a detection material, so that the intrinsic protein amount causing background coloration can be reduced; compared with a method for using the cell as the detection material for carrying out immunofluorescence detection, the detection sensitivity and the specificity are further increased, a detection result is easier and clear to judge, and the AQP4 autoantibodies of a patient can be simply, conveniently and quickly detected.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a human body anti-AQP4 autoantibody detection material and a preparation method thereof. The prepared detection material can be used to detect human body anti-AQP4 autoantibody. Background technique [0002] Human autoantibodies are important serum markers of autoimmune diseases, and their rapid detection has important application value for the diagnosis, treatment and prognosis of autoimmune diseases. With the improvement of the sensitivity and specificity of autoantibody detection, the diagnostic criteria of related autoimmune diseases are also updated and adjusted accordingly. [0003] Neuromyelitis optica (neuromyelitis, NMO) is a serious demyelinating disease of the central nervous system, about 90% of NMO and more than half of patients with aquaporin 4 (aquaporin, AQP4) antibody positive. The discovery of aquaporin 4 antibody in 2004 provided objective evidence for NMO to become a...

AI Technical Summary

Problems solved by technology

However, the main drawbacks of this type of detection method are: (1) Various proteins in the cell may cause background coloring problems

In the specific detection process, although there are idling control cells for comparison, when the expression of exogenous antigens is low, some detection results are still difficult to clearly interpret

(2) The operation process is complicated, and the accuracy of each step directly affects the test results, which is very dependent on the experience of the monitors. In addition, when performing immunofluorescence or flow cytometry experiments, special instruments are required, such as fluorescence microscopes or flow cytometers

The sensitivity and specificity of the IIF method is relatively lower than that of the CBA method, which may be caused by the differences between species in the reaction substrates.

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