Belak, Sandor
- Department of Animal Biosciences, Swedish University of Agricultural Sciences
Abstract
Detection of caliciviruses requires high mutation tolerance and throughput. The development of a rational simple, single tube reverse transcription-real-time quantitative PCR (QPCR) technique for human noroviruses (NV) is reported here. A dual-probe, triple-primer system (NM system) was used for simultaneous detection and preliminary differentiation of NV genogroups in fecal samples. The design was based on a comprehensive analysis of all 1140 NV sequences available in GenBank. A touch-down amplification protocol improved the frequency of detection. The final QPCR was evaluated with 71 fecal samples from outbreak and sporadic cases in Sweden (1997-2004), all calicivirus-positive by electron microscopy. Up to 56 (79%) were positive. The method is more rational than NV detection methods described previously, and should be a developmental basis for large-scale routine methods for detection of NV. (c) 2005 Elsevier B.V. All fights reserved.
Keywords
norovirus; real-time; PCR; QPCR; gastroenteritis; virus quantitation
Published in
Journal of Virological Methods
2006, volume: 132, number: 45659, pages: 69-76
Publisher: ELSEVIER SCIENCE BV
SLU Authors
UKÄ Subject classification
Microbiology in the medical area
Publication identifier
- DOI: https://doi.org/10.1016/j.jviromet.2005.09.006
Permanent link to this page (URI)
https://res.slu.se/id/publ/10088