Human immunodeficiency virus antigen and antibody determination kit and preparation method

Abstract

The invention discloses a human immunodeficiency virus antigen and antibody determination kit obtained through a magnetism particulate immuno chemistry luminescence method. The kit comprises a magnetic separation reagent, a conjugate reagent, a positive reference substance, a negative reference substance and a calibration solution; the magnetic separation reagent is prepared by combining magnetic particles with an envelope antigen and an envelope antibody, and the conjugate reagent is prepared by coupling a detection antigen and a detection antibody with alkaline phosphatase, wherein both the envelope antigen and the detection antigen are recombinant antigens, and both the envelope antibody and the detection antibody are mouse monoclonal antibodies; the calibration solution contains a to-be-detected antigen and a to-be-detected antibody; the magnetic particles have the hydrophobic surfaces and are modified by sulfonyl groups and combined with the antigen or antibody through an adsorption effect. The kit not only has the advantage that little activity of the antigen activity is lost through an envelope method, but also has the advantages that the magnetic particles achieve chemiluminescence and are easy to wash and wide in linear range; the advantages of little antigen loss and short reaction time and the advantages of being easy to wash and wide in linear range are combined for the first time, an inspection result can be consistent with that obtained through an enzyme-immunoassay method, and the reaction time is obviously shortened.

Description

technical field [0001] The invention relates to a magnetic particle chemiluminescence assay kit for human immunodeficiency virus antigen and antibody and a preparation method thereof. Background technique [0002] At present, the human immunodeficiency virus detection kit has been developed to the fourth generation, that is, the antigen and antibody (HIV Ag / Ab) are detected at the same time. This method immobilizes the antigen and antibody in the reaction well of the enzyme-labeled plate by coating, then adds the sample or the processed sample to the reaction well, washes after incubation, and then adds the enzyme-labeled antigen and antibody for reaction after washing. After the reaction is completed, the substrate solution is added for color development, and the negative or positive of the sample can be judged by the depth of the color development. [0003] Generally speaking, enzyme immunoassay has its advantages, that is, the coating of antigen and antibody has little e...

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Problems solved by technology

[0007] Although magnetic particles have the above advantages, they also have disadvantages. The above two types of magnetic beads need to be coupled to antigens or antibodies when used, which means that the antigens or antibodies will have different degrees of activity loss. Due to the high consistency of antibodies in different projects , the coupling sites are all constant regions, and the activity of the variable region (antigen-recognizing region) of the antibody is basically not affected, while the antigens of different projects are highly specific and the active regions are not fixed. If the coupling position is active In this area, errors will occur when the antigen coupled to the magnetic beads binds to the antibody to be tested, resulting in inaccurate test results; not only that, after the antigen is coated or the antibody is coupled to FITC or biotin, it is necessary to add additional magnetic beads for incubation in the detection. steps, resulting in increased detection time

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