Polymerase chain reaction (PCR) and DNA probe-based techniques have improved the sensitivity and specificity that previous diagnostic tests lacked. However, PCR and probing assays are relatively time-consuming and labour intensive, particularly when separate hybridization steps are required to confirm test outcomes.

Recently, real-time PCR technology has greatly improved molecular detection of organisms of veterinary, medical and economic importance. The real-time PCR technology employs similar amplification processes followed in traditional PCR methods (denaturing, annealing and extension) however, the real-time PCR technology allows fast real-time monitoring of a PCR reaction. Amplification and accurate detection of specific targeted nucleic acid sequences is accomplished in a single closed tube, without the need for post-amplification analysis.