An immunosorbent assay for differentiating HIV-1 recent from chronic infection

Abstract

A serological antibody immunosorbent assay for distinguishing HIV-1 recent and chronic infection, fusion expression MP4 containing partial peptides of HIV-1 B, D, CRF01_AE, CRF07_BC, CRF08_BC transmembrane protein As the detection antigen, the specific antibody representing chronic HIV infection in the sample to be tested is captured, and the optical density value of the blue substance produced by the horseradish peroxidase catalyzed substrate TMB is detected to determine the antibody representing chronic HIV infection in the sample to be tested. Presence and content of anti-HIV antibodies. The fusion-expressed tetravalent polypeptide is used as the detection antigen, which can simultaneously detect multiple HIV genotypes, so that the detection range is wider, and the sensitivity and specificity are higher. The classic ELISA indirect method is used for detection, and the operation is relatively simple. It is more useful in the evaluation of control measures and epidemiological surveys of new infection rates in HIV-1 populations.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the establishment of an immunosorbent detection method for distinguishing human immunodeficiency virus type 1 (humanimmunodeficiency virus type 1, HIV-1) from chronic infection, and in high-throughput HIV-1 infection epidemiological investigations and monitoring applications. Background technique [0002] Human immunodeficiency virus (human immunodeficiency virus, HIV) is also known as AIDS virus, and the AIDS caused by it is a chronic fatal infectious disease raging around the world. According to the report of the World Health Organization, as of the end of 2016, there were 36.7 million HIV-infected people in the world. Since the virus was discovered in the past 30 years, it has caused more than 35 million deaths. It seriously endangers public health and is a major public health problem facing the world. [0003] According to the length of time after HIV infection and t...

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Problems solved by technology

However, BED-CEIA has been discontinued due to the synthesis of antigenic materials.

[0009] In addition, these two methods have the following shortcomings in application: (1) Parekh et al. showed that when partial peptides of gp41 of HIV-1B and E subtypes were used as detection antigens, the reaction intensity with blood samples of the same subtype was higher than that of other subtypes. blood type high

The genotypes targeted by existing methods include A, B, C, F, G, H, I, K, AG, AB, AC, BG, AE, D and AD, and the molecular epidemiological survey of HIV in my country shows that my country The main prevalent genotypes are CRF01_AE, CRF07_BC, CRF08_BC and B subtypes. The existing methods are not fully suitable for the detection of new HIV infections in my country and need to be improved and perfected

(2) The existing method based on the recent infection window period and correction coefficient obtained from subtype B blood samples is not completely suitable for the actual situation of multi-subtype epidemics in my country. It is necessary to obtain the corresponding window period in combination with the epidemic background in my country to improve the new infection rate The correction coefficient can accurately calculate the new HIV infection rate in my country

(3) Existing methods have strict limits on the test results of quality control samples, require high technical personnel to operate, and require two experimental operations for primary screening and confirmation, which is not conducive to the prevalence of new HIV-1 infection rates in large-scale populations scientific survey

(4) Existing methods are not clear about gp41 protein as an epitope for recent infection detection, and further research is needed to clarify its possible mechanism of action

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