Figure 1.

One-step qRT-PCR platform vs two-step qRT-PCR in the context of workflow to detect an RNA virus. After sampling the specimen from the patients, RNA materials need to be prepared by extraction and purification (purification of RNAs). The purified RNAs are converted to DNAs by using reverse transcriptase (RT). At this stage, if the patient is infected with RNA viruses, the cDNA fragments derived from the RNA viruses are generated. Then, the virus-originated DNA fragments are sufficiently amplified by the qPCR to a detectable level by the fluorescence signals (amplification/detection). While the one-step qRT-PCR platform can simultaneously achieve both the RT and qPCRs in a single tube, the two-step qRT-PCR needs two separate experimental setups and extra laboratory work and has more chances for contamination by opening the tubes between the RT and PCRs.