Nucleic acid aptamer capable of identifying HCV E1E2 (hepatitis C virus E1E2), nucleic acid aptamer derivatives and screening method and application thereof

Abstract

The invention discloses a nucleic acid aptamer capable of identifying HCV E1E2 (hepatitis C virus E1E2), which is a DNA fragment having a sequence shown in any one of SEQ ID No.1 to SEQ ID No.5. Derivatives of the nucleic acid aptamer include nucleotide-substituted RNA nucleic acid aptamers, skeleton-modified derivatives, peptide nucleic acid derivatives and derivatives of nucleic acid aptamer conjugated with radioactive molecules. A method for screening the nucleic acid aptamer comprises the following steps: firstly preparing HCV E1E2 proteins with histidine tags and then preparing pET200 / D / LacZ proteins with histidine tags, then immobilizing the proteins and designing a random nucleic acid library, and finally screening nucleic acid aptamers. Specifically, the screening step comprises the following steps: pre-treating the DNA library, performing reverse screening, performing forward screening, and repeatedly screening, to obtain the nucleic acid aptamer with strong competitiveness and optimized sequence length. The nucleic acid aptamer and derivatives thereof can be applied to preparing HCV E1E2 protein detection kits or diagnostic kits as well as agents for suppressing HCV infection in liver cells.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a nucleic acid aptamer, a derivative of the nucleic acid aptamer and a screening method and application thereof. Background technique [0002] Hepatitis C virus (HCV) infection is a serious global health problem, with 200 million infected people worldwide. It is estimated that there are 3-4 million new infections each year. About 40 million to 50 million people in my country are infected with HCV, second only to the number of hepatitis B carriers. [0003] Currently, the standard treatment for hepatitis C virus is interferon or interferon combined with ribavirin. Treatment of patients infected with genotypes 2 and 3 has an HCV eradication rate of 75% to 90%, however, about 50% of patients infected with HCV types 1 and 4 remain insensitive. Although the addition of ribavirin can enhance the level of sustained antiviral response, it can also bring about s...

AI Technical Summary

Problems solved by technology

Although the addition of ribavirin can enhance the level of sustained antiviral response, it is also associated with serious side effects (such as hemolytic anemia), which often lead to discontinuation of treatment in 20% of patients

Due to the great sequence diversity of the HCV genome, combined with the side effects of combination therapy and the high cost of treatment, it is urgent to develop new methods to effectively treat liver disease patients infected with different HCV genotypes

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