A review on different types of Real-time PCR methods and its optimization

In the recent years, Real-time PCR technique introduced as a choice method for diagnosis of infectious diseases in many laboratories. During each cycle of the PCR reaction, this technique combines the polymerase chain reaction chemistry with the utilization of fluorescent reporter molecules for monitoring the production of amplification products. Therefore, the set of features including the high sensitivity and specificity, repeatable data and low contamination risk has made the Real-time PCR technology as an attractive alternative to conventional PCR. This technique is often used to quantify the level of gene expression. Since the whole Real-time PCR reaction is performed within a closed tube, the risk of contamination is reduced and eventually prevent false-positive results. The aim of present study was to provide a general overview on different types of Real-time PCR methods, their benefits and applications. Keywords: Molecular diagnosis, Real-time PCR, Polymerase chain reaction, Fluorophor