RT-PCR

RT-PCR reverse transcription polymerase chain reaction (RT-PCR) is highly specific and sensitive molecular method for detecting the presence of specific genetic material. This technique combines reverse transcription that produces cDNA (complementary DNA) from messenger RNA and amplifying specific DNA targets by specific primers using a conventional thermo-cycler reaction also called PCR. This is the most sensitive and gold standard technique, which primarily applied for detection and quantification of gene expression at RNA level. Because of this high sensitivity, the RT-PCR is being increasingly used to quantitative small but biologically pertinent changes in gene expression and detection of RNA viruses (eg SARS-COV2, Influenza viruses).

RT-PCR is carried out as either a single reaction or two-step reaction. In single reaction, the entire reactions from RNA to cDNA and amplification of cDNA are carried out in a test tube. In two-step reaction, reverse transcription that means PCR to cDNA conversion is carried out in a tube and cDNA amplification occur in a separate tube. Using reverse transcription based real-time PCR over other methods to quantify gene expression is more specific and cost-effective. RT-PCR is a sensitive method and apart from RNA virus detection currently this technique is widely used in agriculture and medicine research sectors as well as health and disease diagnosis area. [Md. Mohasin]