Isolating viruses from cultured cells and then using immunofluorescence and molecular biology techniques for viral nucleic acid detection has been successfully used for virus identification. Among them, the common method of using molecular biology technology for virus nucleic acid detection is based on the analysis and identification of polymerase chain reaction (PCR) technology.
A replicon, in the usual sense, is a fragment that starts from a DNA replication origin during DNA replication and is finally completed by the replication fork created from this origin. It contains the control elements needed for replication. The viral replicon refers to the nucleic acid material that guides the replication of the virus, that is, its own DNA or RNA. In virus research, by amplifying and sequencing the sequence of the replicon of a specific virus, the type of virus can be quickly and high-throughput identified.
Using molecular technology to identify the viral genome from clinical samples directly is one of the major discoveries of the 21st century. We have the most advanced virus nucleic acid detection technology.
They are undoubtedly the leading technologies for the rapid detection and identification of most known human viruses. PCR can amplify a specific region of a DNA sequence 106 times in vitro, so it is an extremely sensitive detection method. PCR can also be used for the identification of viral RNA. You only need to reverse transcribe the RNA into DNA and then perform PCR analysis. This method is called reverse transcription PCR (RT-PCR). Since Zhang and Evans first reported it in 1991, there have been reports on the use of RT-PCR to identify influenza viruses.
RT-PCR detects influenza virus is faster than the end-point detection method, and the sensitivity is equal to or better than that of the cell culture method. A previous study on the detection of respiratory syncytial virus from children’s nasal secretions showed that Real-time PCR is the most sensitive quantitative method compared with immunofluorescence and cell culture methods. However, the high mutation rate of the virus will cause drastic changes in the nucleic acid sequence of the virus, resulting in a mismatch with the original PCR primers. Therefore, there is a need to invent a rapid and universal virus detection method. Although the NASBA method is considered to have a high sensitivity, it is not widely used at present due to the difficulty of preparing the NASBA reaction mixture by oneself and the expensive commercial kits. Lawrence Livermore National Laboratory invented a new molecular biology method for microbial detection, which can quickly detect multiple virus types in the same sample. The Lawrence Livermore Microbial Detection Array (LLMDA) method uses probes against various viral genomic DNA to complete virus detection within 24 hours. In addition, these oligonucleotide probes can also detect new species that have homology with known microorganisms.
We combine infection and analytical expertise to provide our clients with the most powerful portfolio of antiviral and antimicrobial in vitro testing services. Facing an increasing demand for new antiviral and antimicrobial compounds for the treatment of infectious diseases, Creative Diagnostics can test these compounds in vitro to determine their potential efficacy in vivo models.