Detection method for slow virus infected cell

Abstract

The invention discloses a detection method for a slow virus infected cell. The method comprises the steps of: synthesizing a single stranded random oligonucleotide library, and infecting an HEK293 cell with MOI (multiplicity of infection)=5; incubating the random oligonucleotide library with a target molecule together for 1h at a temperature of 4DEG C, then washing the monolayer of the cell with PBS (phosphate buffer solution); conducting enzymatic hydrolysis to gain a cell which is resuspended in 0.5ml of H2O at a temperature for 5min so as to release a combined oligonucleotide ligand candidate molecule, generating a new subordinated library through PCR (polymerase chain reaction) or RT-PCR (reverse transcription-polymerase chain reaction), then carrying out combination with the target molecule for a second round screening, repeating the circulation for several times so as to screen out a nucleic acid ligand able to be specifically bound with the target molecule, marking the nucleic acid ligand with Cy5, and incubating the nucleic acid ligand with the slow virus infected HEK293 cell together, performing flow cytometry analysis after washing, cloning the identified nucleic acid ligand to a vector for sequence analysis. The method of the invention can detect a slow virus infected human tissue cell rapidly, simply and effectively, and also can be used for detection of other virus infection or the generation processes of other diseases.

Description

technical field [0001] The invention relates to a biological detection method, and specifically provides a detection method for lentivirus-infected cells. Background technique [0002] Nucleic acid ligand (Aptamer) is an oligonucleotide sequence (DNA or RNA), which is also called nucleic acid antibody because it can bind to the corresponding target molecule with high specificity and high affinity. Nucleic acid ligands are oligonucleotide sequences that are isolated from artificially synthesized single-stranded random oligonucleotide libraries using an in vitro screening and amplification technique, and can specifically bind to target molecules with high efficiency. The in vitro screening process used to obtain ligands is called Systematic Evolution of Ligands by Exponential Enrichment (SELEX). [0003] At present, researchers have screened a variety of ligands that specifically bind to proteins, nucleic acids, oligopeptides, amino acids, organic substances, metal io...

AI Technical Summary

Problems solved by technology

However, the detection of lentivirus-infected cells is cumbersome at present

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