Pseudovirus particle containing human immunodeficiency virus RNA segment and preparation method thereof

Abstract

The invention discloses a preparation method for a pseudovirus particle containing a human immunodeficiency virus RNA segment. The preparation method comprises the following steps: 1) preparing a clone plasmid containing a MS2 gene sequence; 2) screening an HIV gene conserved sequence and preparing an HIV clone plasmid; 3) performing double-plasmid cotransfection to induce the expression of the pseudovirus particle. The pseudovirus particle prepared according to the preparation method is obviously increased, is high in stability, can be stored for a long time, is high in purity, contains high-purity RNA, contains no DNS and is suitable for large-scale production and application.

Description

technical field [0001] The invention relates to human immunodeficiency virus, in particular to a pseudovirion particle containing human immunodeficiency virus RNA fragment and a preparation method thereof. Background technique [0002] AIDS (AIDS) is a major public health problem facing the world, caused by the human immunodeficiency virus (HIV). At the end of 2015, there were an estimated 37 million people living with AIDS worldwide, and more than 18 million (approximately 18.2 million) people living with HIV were currently receiving antiretroviral drug treatment, double the number in 2000. A similar number of people remain without access to treatment, most of whom are unaware of their HIV-positive status. 40% of people living with HIV (over 14 million) remain ignorant of their status. [0003] At present, the quantification of HIV viral load is mostly detected by fluorescent quantitative RT-PCR method. This method has high sensitivity, strong specificity, and wide linear...

AI Technical Summary

Problems solved by technology

The reported HIV commercial products use different gene fragments such as pol, gap, and env to construct pseudoviruses, and the selected length is generally 300bp-500bp. Therefore, the scope of application of the constructed pseudovirions is severely limited, and only the corresponding reagents This is also the main reason why all kinds of quantitative kits cannot share quality control products and quantitative differences

Using conventional pseudovirus construction methods, the size of the inserted foreign fragment is limited

Studies have confirmed that if the packaging efficiency exceeds 500bp, the packaging efficiency will decrease rapidly, which will greatly reduce the yield of the pseudovirus. The sequence of each HIV gene fragment sequence is connected to the plasmid in series, and the size of the packaging fragment exceeds 500bp, which affects the packaging efficiency, that is, the pseudovirus particle. drop in yield

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