Key result
A triplex real-time PCR assay demonstrated equivalent performance to monoplex real-time PCR across a broad dynamic range (10^2 - 10^7 copies/assay) with a limit of detection of 10^2 copies/assay.
Why the study?
Does a triplex real-time PCR assay perform equivalently to monoplex real-time PCR for the detection and quantification of norovirus and sapovirus?
Does a triplex real-time PCR assay perform equivalently to monoplex real-time PCR for the detection and quantification of norovirus and sapovirus?
A newly developed triplex real-time PCR assay provides an efficient and equivalent alternative to monoplex PCR for the simultaneous detection of norovirus and sapovirus.
To improve detection of norovirus (NoVGI, NoVGII) and sapovirus (SaV), a simultaneous quantitative RT-PCR method was established. This triplex real-time PCR method was evaluated using a combination of optimized specific primers and probes. The performance of the developed PCR assay was equivalent to that of monoplex real-time PCR across a broad dynamic range of 10(2) -10(7) copies/assay using plasmid DNA standards. The limit of detection was 10(2) copies/assay. The quantitative value was comparable with that of monoplex real-time PCR of stool samples. Our triplex real-time PCR is useful for detection of NoV and SaV infections.
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Niwa et al. (2013) studied Norovirus and sapovirus infections. Triplex real-time PCR method vs. Monoplex real-time PCR was evaluated on Assay performance and limit of detection. A triplex real-time PCR assay demonstrated equivalent performance to monoplex real-time PCR across a broad dynamic range (10^2 - 10^7 copies/assay) with a limit of detection of 10^2 copies/assay.
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