title: Comparison of RNA extraction methods for the detection of a norovirus surrogate in ready-to-eat foods.
abstract: Four nucleic acid extraction methods were evaluated for the purpose of quantifying a norovirus surrogate (murine norovirus [MNV-1]) concentrated from different food samples. Simple (strawberries and lettuce) and complex (sliced turkey breast, soft-shell clams, and potato salad) food matrices were inoculated with a viral suspension containing high (4×10(5) PFU) or low (4×10(3) PFU) numbers of viral particles. MNV-1 was eluted using either the Pulsifier™ or repetitive pipetting. The four methods were based on using magnetic silica (MiniMAG), non-magnetic silica (bioMérieux Basic kit), silica membrane (Qiagen kit), and phenol (TriReagent) for RNA extraction. The greatest recovery of viral RNA from simple matrices was obtained using magnetic silica for both inoculation levels. For strawberries, the addition of pectinase during the elution step improved RNA recovery when the Pulsifier was used with silica membrane extraction and when repetitive pipetting was used with magnetic silica extraction. In the case of complex matrices, the extraction of high or low numbers of MNV-1 was highest overall using magnetic silica. The exception was soft-shell clams with a high viral load, in which the greatest recovery was obtained with the phenol-based method. In general, magnetic silica was the most effective for extracting both high and low numbers of MNV-1 particles from a wide range of foods.
authors: Institute of Nutraceuticals and Functional Foods (INAF), Université Laval, Québec, Canada.
authors: Maryline
authors: M
authors: Girard
authors: Rocío
authors: R
authors: Morales-Rayas
authors: Julie
authors: J
authors: Jean
mesh_terms: D000818:Animals; D049872:Bivalvia / Q000821:virology; D002460:Cell Line; D057140:Fast Foods / Q000821:virology*; D005506:Food Contamination; D005516:Food Microbiology; D031985:Fragaria / Q000821:virology; D005638:Fruit / Q000821:virology; D018545:Lactuca / Q000821:virology; D008460:Meat / Q000821:virology*; D051379:Mice; D029322:Norovirus / Q000235:genetics* / Q000302:isolation & purification; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012758:Shellfish / Q000821:virology*; D011198:Solanum tuberosum / Q000821:virology; D014422:Turkeys; D014675:Vegetables / Q000821:virology; D019562:Viral Load
chemical_list: D012367:RNA, Viral