After human infection with immunodeficiency virus (HIV), antibodies are generally produced in about 1 month. This period before the appearance of antibodies is called the “window period” of infection. At present, the diagnosis of HIV infection is mainly based on the detection of anti-HIV antibodies in the serum of infected persons, and therefore early detection of those who have not yet produced antibodies often leads to missed detection. In the early stage of HIV infection, the virus replicates in the human body, producing a virus-associated protein, and the p24 antigen protein is a component of the nucleocapsid of the viral structural protein. Early detection can be achieved by detecting the hiv p24 protein in the blood to more effectively control the spread of HIV. The fourth generation of diagnostic reagents has been established internationally, that is, the detection of HIV antibodies and the detection of p24 antigens can detect HIV-infected individuals earlier.
The diagnostic technology for HIV infection has developed rapidly in the past 10 years. In 1998, the fourth generation of diagnostic reagents, namely HIV antigen and antibody reagents, appeared in the world. By detecting the virus’s .p24 protein, it is possible to diagnose AIDS early, control its rapid spread, and assist in the diagnosis of disease development and evaluation of antiviral treatment effects. In this experiment, we have obtained a large number of anti-HIV-p24 polyclonal and multiple monoclonal antibodies, using double antibody sandwich method and indirect ELISA antibody amplification system to detect HIV-p24 protein, and compared different coated antibodies and labeled antibodies. And the amplification system, preferably a sensitive and specific detection method.
It is argued that the conjugate vaccine of HIV-infected patients is several times that of asymptomatic patients. The acute phase is the best time to start HIV infection treatment, but there are often no clinical symptoms at this time, so the laboratory that provides early diagnosis basis is provided. Detection methods are especially important. At present, the sensitivity of detection of four generations of enzyme-linked reagents that cannot be distinguished by antigens and antibodies in the market has been improved compared with the past. However, it has been reported in the literature that since antigens and antibodies are simultaneously coated on the reaction plate, there is a possibility of mutual interference, which affects immunity. There is a gap between the specificity of the reaction and the sensitivity of the reagent capable of detecting the p24 antigen alone, and the detection time of the acute infection specimen is significantly delayed. In this study, the reagents that can distinguish HIV-1p24 antigen from antibodies are not only provided for HIV infection, especially the detection of HIV-1p24 antigen positive provides a powerful early diagnosis of acute infection. stand by. In this study, in silico vaccine design HIV-1 p24 antigen-positive cases alone, 4 of which were negative by four-generation enzyme-linked reagents that could not distinguish antigen-antibodies, and one of the 8 cases of antigen-antibody-positive ones were negative. It is suggested that in clinical tests, for high-risk behaviors or suspected cases, high-sensitivity four-generation reagents for detecting antigens and antibodies should be used as much as possible to avoid positive missed detection.
No p24 protein was detected in 22 HIV antibody-positive sera using the double antibody sandwich method or the immunoamplification system. The reason may be that the p24 antibody is produced in the body of the infected person, but it has not entered the onset stage, and the amount of virus is small. On the other hand, it is possible that the immune complex formed by the p24 antigen and the antibody in the serum is not present in this experiment. Being effectively dissociated. Under laboratory conditions, the antigen suspension containing HIV-p24 protein was serially diluted with normal human serum. After lysing, the p24 protein was examined by double antibody sandwich method to simulate the virus present in the serum of the infected person, and the sensitivity of the test was determined. And the amount of virus. Due to the limited conditions, this study used the imported detection p24 protein antibody vaccine reagent as a control and obtained similar sensitivity.
The test results show similar sensitivity to similar products in the world, and can be extended to clinical specimen testing, thus providing a simple, rapid and economical method for the evaluation of window-infected patients and clinical efficacy evaluation. Diagnostic reagents provide a basis and antibody library.