Murine cytomegalovirus real-time fluorescent quantitative PCR (Polymerase Chain Reaction) detection method

Abstract

The invention relates to a murine cytomegalovirus real-time fluorescent quantitative PCR detection method comprising the following steps of: standard plasmid construction, specific primer and fluorescent probe design, real-time fluorescent quantitative PCR amplification and detection standard curve establishment, RNA extraction of an infected virus sample, cDNA preparation and validity verification and result detection judgment, wherein the sequence of a forward primer of a specific primer is shown as follows: 5'-AGCGTCGATGCAGGGGCTTT-3', and the sequence of a reverse primer thereof is shown as follows: 5'-CGGCGGAACGGGGGCTGGTA-3'; and the sequence of a fluorescent probe is shown as follows: 5'-CGGGGGCGTTCCGAAAACGA-3'. The invention has the advantages of high detection sensitivity, short detection time, good specificity, sensibility, repeatability and stability, can be simultaneously used for carrying out sample analysis on a large scale and is suitable for the early diagnosis of virus infection.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a virus detection method, in particular to a mouse cytomegalovirus real-time fluorescent quantitative PCR detection method. Background technique [0002] Mouse cytomegalovirus (Murine Cytomegalovirus, MCMV) belongs to the Herpesviridae family and is a virus that can cause persistent infection. When the body's resistance is low, it can cause acute disease and death, leading to the failure of the experiment. [0003] At present, there are many methods for detecting various viruses, such as serological tests, tissue block culture and co-cultivation techniques, nucleic acid hybridization and conventional PCR detection techniques, among which the sensitivity, specificity, stability and operability of conventional PCR detection techniques are all high. It is better, but there is cross-contamination in the treatment after the conventional PCR reaction, and it is relatively easy to have false...

AI Technical Summary

Problems solved by technology

[0003] At present, there are many methods for detecting various viruses, such as serological tests, tissue block culture and co-cultivation techniques, nucleic acid hybridization and conventional PCR detection techniques, among which the sensitivity, specificity, stability and operability of conventional PCR detection techniques are all high. It is better, but there is cross-contamination after the conventional PCR reaction, and it is more prone to false positive results, and the experiment time is relatively long

Images