Novel light-upon-extension real-time PCR assays for detection and quantification of genogroup I and II noroviruses in clinical specimens

Nordgren, Johan, 1980- (author): Linköpings universitet,Molekylär virologi,Hälsouniversitetet

Bucardo, Filemón (author): Department of Microbiology, University of León, León, Nicaragua

Dienus, Olaf (author): Microbiological Laboratory, Ryhov County Hospital, Jönköping, Sweden

Svensson, Lennart (author): Linköpings universitet,Molekylär virologi,Hälsouniversitetet

Lindgren, Per-Eric (author): Linköpings universitet,Medicinsk mikrobiologi,Hälsouniversitetet

(creator_code:org_t)

2008
2008
English.
In: Journal of Clinical Microbiology. - 0095-1137 .- 1098-660X. ; 46:1, s. 164-170

Related links:: http://urn.kb.se/res...; show more...; https://urn.kb.se/re...; https://doi.org/10.1...; show less...

Abstract Subject headings

Norovirus is now recognized as the leading cause of nonbacterial acute gastroenteritis in adults, causing numerous outbreaks worldwide. We have developed two novel light-upon-extension (LUX) real-time PCR assays for detection and quantification of norovirus genogroups I and II. The LUX system uses a fluorophore attached to one primer having a self-quenching hairpin structure, making it cost-effective and specific. The assays were evaluated against clinical stool specimens (n = 103) from Sweden and Nicaragua and compared to established methods. The norovirus assay detected more positive stool specimens (47/103) than conventional PCR (39/103) and corresponded to a TaqMan real-time PCR, with the exception of one specimen. Furthermore, the assays correctly identified all (n = 11) coded control specimens in a reference panel containing various genogroups and genotypes. Both LUX real-time PCR assays had a wide dynamic range, detecting from < or = 10(1) to 10(7) genes per reaction, resulting in a theoretical lower limit of < or = approximately 20,000 viruses per gram of stool. No cross-reactivity was noticed with specimens containing other enteric viruses, and by using melting curve analysis we could differentiate between norovirus genogroups I and II.

By the author/editor: Nordgren, Johan, ...; Bucardo, Filemón; Dienus, Olaf; Svensson, Lennar ...; Lindgren, Per-Er ...

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