A non-diagnostic HIV antibody immunoassay method and kit

Abstract

The invention relates to a non-diagnosis-targeted HIV antibody immunity detecting method and a kit. The method comprises the following steps: specifically capturing and detecting an HIV antibody in a sample by using an HIV antigen, then adding a biotinylated secondary antibody, and incubating to form an antigen-antibody-biotinylated secondary antibody compound; adding a biotinylation primer Biotin-I in the presence of streptavidin, incubating to combine the compound with the Biotin-I, and then adding biotinylated hairpin chain Biotin-H1 and hairpin chain H2 to carry out hybridization reaction; adding avidin marked horseradish peroxidase, catalyzing a substrate solution to develop color, and carrying out quantitative detection on the HIV antibody by determining absorbance. HCR is guided into ELISA, signals are amplified by the HCR, ultra-sensitive detection of the HIV antibody is realized, and lower limit of detection is as low as 9.5 pg / mL, and is higher than that of the traditional ELISA by about 2 orders of magnitude.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, in particular to a non-diagnostic HIV antibody detection method and kit. Background technique [0002] Human immunodeficiency virus (Human immunodeficiency virus, HIV) is the pathogen that causes acquired immunodeficiency syndrome (Acquired Immune Deficiency Syndrome, AIDS). HIV antibody is produced by the human immune response after the HIV virus enters the human body. This antibody is basically ineffective against the HIV virus, but it can reflect the HIV infection status to a certain extent and is a physiological indicator of the human body. [0003] Detecting HIV antibodies in blood is currently the most commonly used laboratory method for detecting HIV infection. Generally, it needs to go through two steps: first, do a preliminary screening test, and if it is positive, then do a confirmatory test. Only when the confirmation test is positive can the diagnosis of AIDS be made. Viral infe...

AI Technical Summary

Problems solved by technology

Methods such as GPAT and dot immune colloidal gold rapid test are subjective, expensive, and difficult to store.

The disadvantage of CLIA is that the luminescent reaction of labeled catalytic enzymes or chemiluminescent molecules is unstable, which is intermittent and flashing, and it is prone to fission during the reaction process, resulting in unstable reaction results.

In addition, the binding phase and the free phase need to be separated during detection, which requires many steps and high testing costs.

ELISA is still the most commonly used clinical method for detecting HIV antibodies. It has the advantages of sensitivity, specificity, and rapidity. However, the detection limit of ELISA is still limited and cannot meet the detection of low-concentration HIV antibody levels in the early stage of infection.

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