Recently, TaqMan technology has combined the 5 nuclease activity of the Taq DNA polymerase and forster resonance energy transfer to detect and quantify amplification product in a closed tube format. Using this technology real-time PCR has been developed to detect the nucleic acid [164, 165] . This is most sensitive and rapid method to detect the nucleic acid. The viral RNA can be consistently detected over a seven log range, the lowest of which corresponded to as few as 10-100 RNA per volume tested. The test can be performed in 2 hours or less on a portable instrument and sample can be held at ambient temperatures. Real-Time chemistry allows for the detection of PCR amplification during the early phases of the reaction and as the name suggest, realtime PCR monitors the progress of a PCR reaction in the real-time. At the same time, a relatively small amount of PCR products (DNA, CDNA, or RNA) can be quantified. Measuring the kinetics of the reaction in the early phases of PCR provides a distinct advantage over traditional PCR detection. Traditional methods use agarose gels for detection of PCR amplification at the final phase or end point of the PCR reaction which is associated with considerable hands on time, posses a serious hazard for amplification product Veterinary Medicine International 7 carryover, and limits the number of specimen that can be processed simultaneously. It is hard to differentiate the 5 fold changes on gel whereas real-time PCR is able to detect even twofold changes (e.g., 10 versus 20 copies). In addition to the widely exploited 5 -nuclease (TaqMan) system using duallabelled probes [39, 161, 166] and modified MGB probes [167] , assays have also been developed using other rRT-PCR formats such as those using hybridization probes [146] and PriProET [168] .