Virus-like particle containing RNA virus nucleic acid and its preparing method and use

Abstract

The present invention belongs to the field of virus gene engineering, and relates to virus-like particle containing RNA virus nucleic acid and its preparation process and application. The virus-like particle is RNase resistant, and contains coat protein and inside recombinant gene sequence including partial MS2 bactereophage sequence, RNA virus nucleic acid sequence and partial RNA virus antigen. The virus-like particle of the present invention may be used as stable and reliable RNA quality controlling and standard article without biological infectious danger. The present invention provides also RT-PCR reagent kit amplification segment suitable for detecting virus RNA in several methods. The virus-like particle containing partial RNA virus antigen may be used as the mold for research of the interaction between virus and host cell, as antisense RNA tool for directional transportation of small molecule medicine, and in vaccine research.

Description

【Technical field】 [0001] The invention relates to the field of virus genetic engineering, in particular to a virus-like particle containing RNA virus nucleic acid and a preparation method and application thereof. 【Background technique】 [0002] RNA viruses such as hepatitis C virus (HCV), human immunodeficiency virus (HIV), and severe acute respiratory syndrome coronavirus (SARS-CoV) are a class of RNA viruses that can cause severe The pathogens of diseases, at present, there are mainly two types of methods for the detection of these pathogens (Collins ML, et al., Nucleic Acids Res.1997, 25 (15): 2979-84; MulderJ, et al., J Clin Microbiol.1994 , 32 (2): 292-300), one is antigen-antibody-based immunoassay methods, which mainly detect virus antigens in samples or antibodies produced by the body. The other is the use of nucleic acid to detect viral RNA in patient specimens. Immunological detection is a routine method for diagnosing viruses. However, compared with nucleic acid...

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Problems solved by technology

Human error will cause the loss of sample nucleic acid, reduce the amount of target nucleic acid and lead to false negative results

(3) Influencing factors of the reaction system: low enzyme activity during reverse transcription, degradation of the target fragment due to unsuitable storage conditions, unreasonable primer design, and unsatisfactory thermal cycle process can also lead to Inaccurate test results

But the target sequence used for HIV-1 quality control in the virus-like particle expressed by this expression vector is too small, is not suitable for multiple methods or the application that contains the RT-PCR kit of multiple detection virus RNA; Simultaneously virus-like particle is not very Stable, and unable to produce virus-like particles with HIV-1 partial antigens, its application is small

[0009] In summary, in the RT-PCR detection of RNA viruses, due to many factors affecting the experimental process, such as the random error in the measurement operation of the experimenter, the temperature difference between the wells of the amplification instrument, and the concentration of inhibitors after the nucleic acid extraction of the sample. Residues, the concentration of the target nucleic acid to be amplified, the efficiency of reverse transcription, and reagent problems, etc., these factors may affect the efficiency of PCR amplification, thereby causing deviations in results

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