PCR primer set, probe and its kit and detection method for detecting human immunodeficiency virus type 1

Abstract

The invention discloses PCR primer groups and a probe used for detecting human immunodeficiency virus type I, a kit containing the PCR primer groups and the probe, and a detection method. The kit contains the high sensitivity primer groups and the probe. According to the detection method, multiple circulation is designed, and temperature is increased to a primer annealing temperature, and extremely high sensitivity and specificity are achieved. According to the minimum detection limit, in each time of detection, 2 copies HIV-1 DNA / millions of cells can be realized, and sensitivity of the detection method is higher than that of other detection methods. A cell quantitative system is introduced into the detection method, so that quantification of HIV-1 DNA and cell number can be realized simultaneously in a same reaction. A quantitative standard substance is added into the kit so as to solve a problem of HIV DNA auantitative traceability in the prior art, wherein the quantitative standard substance is simple to prepare, is stable, and is reliable in traceability. The PCR primer groups, the probe, the kit, and the detection method can be used for HIV infection early detection, HIV infection screening of infants or high risk groups, HIV-1 virus repository detection, antiviral drug therapeutic effect evaluation, disease relapse control, and radical cure standard establishment; operation is simple; cost is low; clinical application prospect is promising; and it is beneficial for popularization and application.

Description

technical field [0001] The invention belongs to the field of virus detection, and relates to a PCR primer set, a probe, a kit thereof and a detection method for detecting human immunodeficiency virus type 1. Background technique [0002] AIDS is acquired immunodeficiency syndrome (acquired immunodeficiency syndrome, AIDS), is a chronic fatal infectious disease, caused by human immunodeficiency virus (human immunodeficiency virus, HIV), divided into human immunodeficiency virus type 1 (HIV- 1) and human immunodeficiency virus type 2 (HIV-2), HIV-1 is currently the main strain circulating in the world including China, and HIV-2 is currently only circulating in West Africa. After HIV infection, the immune function of the human body is deficient, and a series of clinical syndromes such as opportunistic infections and tumors occur, and the fatality rate is almost 100%. According to 2011 data, AIDS has spread to more than 200 countries and regions around the world, the total numb...

AI Technical Summary

Problems solved by technology

This method requires high personnel and equipment, is expensive, requires a large amount of samples (120-180mL blood), is time-consuming and laborious, and there are many factors that affect the accuracy of the results, the detection rate is low, and it is not suitable for tissue biopsy

②CD4+T lymphocyte count is mostly detected by CD4 / 8 / 3 flow cytometry, but there is no evidence that it can directly reflect changes in the HIV virus reservoir, and whether there is a certain correlation needs to be further verified

[0010] In summary, traditional HIV detection methods are limited by the characteristics of "window period", time-consuming, costly, high specimen requirements, and low detection sensitivity. It is urgent to develop a highly sensitive HIV DNA detection method

At present, there are few HIV DNA detection kits at home and abroad, and the sensitivity of the HIV DNA PCR detection method reported in the article is 10 copies or more of HIV DNA per million cells.

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