Real-time PCR

Compared with conventional PCR, and in particular compared with nested PCR, real-time PCR has the advantage that the amplification process is recorded in real time. The risk of contamination is very much reduced since there is no need to open the reaction tube once the amplification has started, i.e. the post-amplification detection step is omitted. Therefore, and because more of the process can be automated, it takes much less time and labour to do real-time PCR than nested PCR.

For the same reasons, the risk of sample switching is lower in realtime PCR. The sensitivity is comparable between real-time PCR and nested PCR. Real-time PCR results are digitised instantly and analysed using standardised computer algorithms, thus minimising human errors, and may provide test results within 1 h after the start of the analysis, whereas analysis based on nested PCR may take hours to complete.