title: Comparison of three extraction methods to detect noroviruses in dairy products.
abstract: Noroviruses (NoV) are currently the most common cause of viral foodborne diseases and RT-qPCR is widely used for their detection in food because of its sensitivity, specificity and rapidity. The ISO/TS (15216-1, 15216-2) procedures for detecting NoV and HAV in high-risk food categories such as shellfish, bottled water and vegetables were published in 2013. Milk products are less implicated in foodborne viral outbreaks but they can be contaminated with fruit added to these products or by the food handler. Thus, the development of sensitive and reliable techniques for the detection of NoV in dairy products is needed to ensure the safety of these products. The aim of this study was to develop a RT-qPCR based method for the detection of NoV in milk products. Three methods were tested to recover NoV from artificially contaminated milk and cottage cheese. The selected method was based on the use of proteinase K and the recovery efficiencies ranged from 54.87% to 98.87% for NoV GI, 61.16%-96.50% for NoV GII. Murine norovirus and mengovirus were used as process controls and their recovery efficiencies were respectively 60.59% and 79.23%. The described method could be applied for detecting NoV in milk products for routine diagnosis needs.
authors.affiliation: Université Paris-Est, Anses, Laboratory for Food Safety, 14 rue Pierre et Marie Curie, F-94700, Maisons-Alfort, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Hennechart-Collette
authors.affiliation: Université Paris-Est, Anses, Laboratory for Food Safety, 14 rue Pierre et Marie Curie, F-94700, Maisons-Alfort, France.
authors.forename: Sandra
authors.initials: S
authors.lastname: Martin-Latil
authors.affiliation: Université Paris-Est, Anses, Laboratory for Food Safety, 14 rue Pierre et Marie Curie, F-94700, Maisons-Alfort, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Fraisse
authors.affiliation: Université Paris-Est, Anses, Laboratory for Food Safety, 14 rue Pierre et Marie Curie, F-94700, Maisons-Alfort, France. Electronic address: sylvie.perelle@anses.fr.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Perelle
mesh_terms: D000818:Animals; D002611:Cheese / Q000382:microbiology*; D019286:Endopeptidase K; D005506:Food Contamination / Q000032:analysis; D005516:Food Microbiology; D016679:Genome, Viral; D057230:Limit of Detection; D008574:Mengovirus / Q000235:genetics / Q000302:isolation & purification; D051379:Mice; D008892:Milk / Q000821:virology*; D029322:Norovirus / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D014773:Virology / Q000379:methods*
chemical_list: D012367:RNA, Viral; D019286:Endopeptidase K