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Development and evaluation of novel one-step TaqMan realtime RT-PCR assays for the detection and direct genotyping of genogroup I and II noroviruses. / Schultz, Anna Charlotte; Vega, Everado; Dalsgaard, Anders; Christensen, Laurids Siig; Nørrung, Birgit; Hoorfar, Jeffrey; Vinjé, Jan.

In: Journal of Clinical Virology, Vol. 50, No. 3, 2011, p. 230-234.

Publication: Research - peer-reviewJournal article – Annual report year: 2010

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Schultz, Anna Charlotte; Vega, Everado; Dalsgaard, Anders; Christensen, Laurids Siig; Nørrung, Birgit; Hoorfar, Jeffrey; Vinjé, Jan / Development and evaluation of novel one-step TaqMan realtime RT-PCR assays for the detection and direct genotyping of genogroup I and II noroviruses.

In: Journal of Clinical Virology, Vol. 50, No. 3, 2011, p. 230-234.

Publication: Research - peer-reviewJournal article – Annual report year: 2010

Bibtex

@article{3d064d0575684d6895c3f1d6ba0ebb8e,
title = "Development and evaluation of novel one-step TaqMan realtime RT-PCR assays for the detection and direct genotyping of genogroup I and II noroviruses",
publisher = "Elsevier BV",
author = "Schultz, {Anna Charlotte} and Everado Vega and Anders Dalsgaard and Christensen, {Laurids Siig} and Birgit Nørrung and Jeffrey Hoorfar and Jan Vinjé",
year = "2011",
doi = "10.1016/j.jcv.2010.12.001",
volume = "50",
number = "3",
pages = "230--234",
journal = "Journal of Clinical Virology",
issn = "1386-6532",

}

RIS

TY - JOUR

T1 - Development and evaluation of novel one-step TaqMan realtime RT-PCR assays for the detection and direct genotyping of genogroup I and II noroviruses

A1 - Schultz,Anna Charlotte

A1 - Vega,Everado

A1 - Dalsgaard,Anders

A1 - Christensen,Laurids Siig

A1 - Nørrung,Birgit

A1 - Hoorfar,Jeffrey

A1 - Vinjé,Jan

AU - Schultz,Anna Charlotte

AU - Vega,Everado

AU - Dalsgaard,Anders

AU - Christensen,Laurids Siig

AU - Nørrung,Birgit

AU - Hoorfar,Jeffrey

AU - Vinjé,Jan

PB - Elsevier BV

PY - 2011

Y1 - 2011

N2 - BackgroundCurrent detection and genotyping methods of genogroup (G) I and II noroviruses (NoVs) consist of a 2-step approach including detection of viral RNA by TaqMan realtime RT-PCR (RT-qPCR) followed by conventional RT-PCR and sequencing of partial regions of ORF1 or ORF2. ObjectiveTo develop novel long-template one-step TaqMan assays (L-RT-qPCR) for the rapid detection and direct genotyping of GI and GII NoVs and to evaluate the sensitivity and specificity of the assays. Study designGI and GII-specific broadly reactive L-RT-qPCR assays were developed by combining existing NoV primers and probes targeting the open reading frame (ORF)1–ORF2 junction as well as region C at the 5′–ORF2. The assays were validated using GI and GII RNA transcripts and a coded panel of 75 stool samples containing NoV strains representing 9 GI genotypes and 12 GII genotypes, as well as sapoviruses, astroviruses, polioviruses, and rotaviruses. L-RT-qPCR products were typed by sequencing. ResultsThe novel GI and GII L-RT-qPCR assays detected and typed all but one of the NoV positive panel samples. As few as 5–500 RNA copies could be accurately typed by sequencing of amplicons. ConclusionsWe developed novel one-step TaqMan RT-qPCR assays for the sensitive detection and direct genotyping of GI and GII NoVs from clinical and environmental matrices

AB - BackgroundCurrent detection and genotyping methods of genogroup (G) I and II noroviruses (NoVs) consist of a 2-step approach including detection of viral RNA by TaqMan realtime RT-PCR (RT-qPCR) followed by conventional RT-PCR and sequencing of partial regions of ORF1 or ORF2. ObjectiveTo develop novel long-template one-step TaqMan assays (L-RT-qPCR) for the rapid detection and direct genotyping of GI and GII NoVs and to evaluate the sensitivity and specificity of the assays. Study designGI and GII-specific broadly reactive L-RT-qPCR assays were developed by combining existing NoV primers and probes targeting the open reading frame (ORF)1–ORF2 junction as well as region C at the 5′–ORF2. The assays were validated using GI and GII RNA transcripts and a coded panel of 75 stool samples containing NoV strains representing 9 GI genotypes and 12 GII genotypes, as well as sapoviruses, astroviruses, polioviruses, and rotaviruses. L-RT-qPCR products were typed by sequencing. ResultsThe novel GI and GII L-RT-qPCR assays detected and typed all but one of the NoV positive panel samples. As few as 5–500 RNA copies could be accurately typed by sequencing of amplicons. ConclusionsWe developed novel one-step TaqMan RT-qPCR assays for the sensitive detection and direct genotyping of GI and GII NoVs from clinical and environmental matrices

UR - https://www-ncbi-nlm-nih-gov.globalproxy.cvt.dk/pubmed/21195660

U2 - 10.1016/j.jcv.2010.12.001

DO - 10.1016/j.jcv.2010.12.001

JO - Journal of Clinical Virology

JF - Journal of Clinical Virology

SN - 1386-6532

IS - 3

VL - 50

SP - 230

EP - 234

ER -