Xia, Hongyan
- Department of Animal Biosciences, Swedish University of Agricultural Sciences
- Uppsala University
Abstract
The high sequence variation of RNA viruses necessitates use of degenerate primers and probes or multiple primers and probes in molecular diagnostic assays. We showed previously that PCR amplification in two rounds, first with long target-specific primers and then with short generic primers, followed by detection using long probes, can tolerate sequence variation. Here we demonstrate that long primers and probes of up to 56 nucleotides can also be applied in real-time PCR for the detection of norovirus genogroup II with improved sensitivity. Probe design (method of incorporating quenchers, use of Zen internal quencher or traditional quenchers) greatly affects the sensitivity of the real-time PCR assays.
Keywords
real-time PCR; variation-tolerant capture multiplex assay; norovirus
Published in
Biotechniques
2016, volume: 60, number: 1, pages: 28-34
SLU Authors
Ottoson, Jakob
- Department of Animal Biosciences, Swedish University of Agricultural Sciences
- National Food Agency
- Department of Animal Biosciences, Swedish University of Agricultural Sciences
UKÄ Subject classification
Biomedical Laboratory Science/Technology
Publication identifier
- DOI: https://doi.org/10.2144/000114370
Permanent link to this page (URI)
https://res.slu.se/id/publ/69248