BACKGROUND: Reverse transcription (RT)-PCR for norovirus detection is prone to false-negative results due to inhibitory substances in faeces. An internal control is needed to monitor extraction efficiency and to detect inhibition. OBJECTIVES: To further develop a one-step RT-PCR assay for norovirus detection/genogrouping by addition of MS2 bacteriophage as an internal control. STUDY DESIGN: Our norovirus RT-PCR assay was modified by addition of MS2 phage to the extraction tray and primers/probe for MS2 detection to the reaction mix. The effect of addition of MS2 phage and MS2 primers/probe on the sensitivity/specificity of the PCR assay was examined. RESULTS: The addition of MS2 as an internal control showed no loss of sensitivity or specificity for norovirus detection. CONCLUSIONS: A triplex, one-step, type-specific, real-time RT-PCR with MS2 internal control has been developed for use in routine laboratory diagnosis of norovirus infection.
BACKGROUND: Reverse transcription (RT)-PCR for norovirus detection is prone to false-negative results due to inhibitory substances in faeces. An internal control is needed to monitor extraction efficiency and to detect inhibition. OBJECTIVES: To further develop a one-step RT-PCR assay for norovirus detection/genogrouping by addition of MS2 bacteriophage as an internal control. STUDY DESIGN: Our norovirus RT-PCR assay was modified by addition of MS2 phage to the extraction tray and primers/probe for MS2 detection to the reaction mix. The effect of addition of MS2 phage and MS2 primers/probe on the sensitivity/specificity of the PCR assay was examined. RESULTS: The addition of MS2 as an internal control showed no loss of sensitivity or specificity for norovirus detection. CONCLUSIONS: A triplex, one-step, type-specific, real-time RT-PCR with MS2 internal control has been developed for use in routine laboratory diagnosis of norovirus infection.
Authors: Jennifer L Cannon; Leslie Barclay; Nikail R Collins; Mary E Wikswo; Christina J Castro; Laura Cristal Magaña; Nicole Gregoricus; Rachel L Marine; Preeti Chhabra; Jan Vinjé Journal: J Clin Microbiol Date: 2017-05-10 Impact factor: 5.948
Authors: David M Goldfarb; Andrew P Steenhoff; Jeffrey M Pernica; Sylvia Chong; Kathy Luinstra; Margaret Mokomane; Loeto Mazhani; Isaac Quaye; Irene Goercke; James Mahony; Marek Smieja Journal: J Clin Microbiol Date: 2014-08-27 Impact factor: 5.948
Authors: Jeremy M Woodward; Effrossyni Gkrania-Klotsas; Anthony Yk Cordero-Ng; Aloysious Aravinthan; Betty N Bandoh; Hongxiang Liu; Susan Davies; Hongyi Zhang; Philip Stevenson; Martin D Curran; Dinakantha Kumararatne Journal: Am J Gastroenterol Date: 2015-01-27 Impact factor: 10.864
Authors: John R Ticehurst; Nora Pisanic; Michael S Forman; Carly Ordak; Christopher D Heaney; Edgar Ong; Jeffrey M Linnen; Paul M Ness; Nan Guo; Hua Shan; Kenrad E Nelson Journal: Transfusion Date: 2019-01-31 Impact factor: 3.157
Authors: Jane S Greatorex; Rosanna F Page; Martin D Curran; Paul Digard; Joanne E Enstone; Tim Wreghitt; Penny P Powell; Darren W Sexton; Roberto Vivancos; Jonathan S Nguyen-Van-Tam Journal: PLoS One Date: 2010-02-01 Impact factor: 3.240