Homogeneous immunodetection kit for detecting anti-cyclic citrullinated peptide antibody and application thereof
An anti-cyclic citrullinated peptide and immunodetection technology, applied in the field of immunoassay, can solve the problems of complicated operation, poor specificity, low sensitivity, etc., and achieve the effects of low technical requirements, simple operation and high sensitivity
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[0063] Ⅱ. Implementation plan
[0064] As mentioned earlier, the current research or commercial anti-citrulline antibody kits use indirect detection methods, especially indirect detection by ELISA. Anti-citrulline antibody types in the human body include IgG, IgA and IgM. The traditional indirect reaction mode can only detect IgG antibodies. The indirect detection mode of ELISA can not only detect IgG type antibodies, but also has low sensitivity, poor specificity, and cumbersome operation. To this end, the inventors have conducted a large number of theoretical and experimental studies on the anti-citrulline antibody detection kit.
[0065] The inventors have discovered through research that the antigen obtained by the preparation method of the present invention can be specifically recognized and bound by the anti-citrulline antibody to be tested, forming a sandwich (solid-phased antigen-antibody-label antigen) immune complex It can complete the determination of all types of anti...
Example Embodiment
[0096] According to some specific embodiments of the present invention, the method includes:
[0097] Step R1, mixing the sample to be tested with the first composition and the second composition and then reacting to obtain the first mixture;
[0098] Step R2, mixing the first mixture with the third composition and then reacting to obtain a second mixture;
[0099] Step R3, contacting the energy or active compound with the second mixture to excite the donor to generate singlet oxygen, and the acceptor can react with the received singlet oxygen to generate a detectable chemiluminescence signal;
[0100] Step R4, detecting the presence and / or intensity of the chemiluminescent signal in step R3, so as to determine whether there is an anti-cyclic citrullinated peptide antibody in the test sample and / or determine the content of the anti-cyclic citrullinated peptide antibody.
Example Embodiment
[0101] According to some embodiments of the present invention, the method further includes the step of preparing a standard working curve of the anti-cyclic citrullinated peptide antibody before step R1.
[0102] In some specific embodiments of the present invention, in step R4, the intensity of the chemiluminescence signal in step R3 is detected, and the anti-cyclic citrullinated peptide antibody standard working curve is used to determine the anti-cyclic citrullinated peptide in the sample to be tested. The content of amino acid peptide antibody.
[0103] In the present invention, there is no particular restriction on the manner or order of mixing the sample to be tested with the first composition and the second composition in step R1. For example, in some embodiments of the present invention, in step R1, The sample is mixed with the first composition and the second composition at the same time and then reacted to obtain the first mixture; and in other embodiments of the present ...
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