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Author (up) Afolayan, O.T.; Webb, C.C.; Cannon, J.L. doi  openurl
  Title Evaluation of a Porcine Gastric Mucin and RNase A Assay for the Discrimination of Infectious and Non-infectious GI.1 and GII.4 Norovirus Following Thermal, Ethanol, or Levulinic Acid Plus Sodium Dodecyl Sulfate Treatments Type Research Support, U.S. Gov't, Non-P.H.S.
  Year 2016 Publication Food and environmental virology Abbreviated Journal Food Environ Virol  
  Volume 8 Issue 1 Pages 70-78  
  Corporate Author Thesis  
  Address Department of Food Science and Technology, Center for Food Safety, University of Georgia, 1109 Experiment Street, Griffin, GA, 30223, USA. jcannon@uga.edu  
  Keywords Animals; Biological Assay/*methods; Caliciviridae Infections/*virology; Ethanol/*pharmacology; Gastric Mucins/*metabolism; Hot Temperature; Humans; Levulinic Acids/*pharmacology; Norovirus/chemistry/classification/drug effects/*isolation & purification; Ribonuclease, Pancreatic/analysis; Sodium Dodecyl Sulfate/*pharmacology; Swine; Ethanol; Levulinic acid; Norovirus; Porcine gastric mucin; Sodium dodecyl sulfate; Surrogate; Thermal inactivation  
  Abstract Human noroviruses (NoVs) are a major source of foodborne illnesses worldwide. Since human NoVs cannot be cultured in vitro, methods that discriminate infectious from non-infectious NoVs are needed. The purpose of this study was to evaluate binding of NoV genotypes GI.1 and GII.4 to histo-blood group antigens expressed in porcine gastric mucin (PGM) as a surrogate for detecting infectious virus following thermal (99 degrees C/5 min), 70% ethanol or 0.5% levulinic acid (LV) plus 0.01 or 0.1% sodium dodecyl sulfate (SDS) sanitizer treatments and to determine the limit of detection of GI.1 and GII.4 binding to PGM. Treated and control virus samples were applied to 96-well plates coated with 1 microg/ml PGM followed by RNase A (5 ng/microl) treatment for degradation of exposed RNA. Average log genome copies per ml (gc/ml) reductions and relative differences (RD) in quantification cycle (Cq) values after thermal treatment were 1.77/5.62 and 1.71/7.25 (RNase A) and 1.73/5.50 and 1.56/6.58 (no RNase A) for GI.1 and GII.4, respectively. Treatment of NoVs with 70% EtOH resulted in 0.05/0.16 (GI.1) and 3.54/10.19 (GII.4) log reductions in gc/ml and average RD in Cq value, respectively. LV (0.5%) combined with 0.1 % SDS provided a greater decrease of GI.1 and GII.4 NoVs with 8.97 and 8.13 average RD in Cq values obtained, respectively than 0.5% LV/0.01 % SDS. Virus recovery after PGM binding was variable with GII.4 > GI.1. PGM binding is a promising surrogate for identifying infectious and non-infectious NoVs after capsid destruction, however, results vary depending on virus strain and inactivation method.  
  Publisher Place of Publication Editor  
  Language English Summary Language Original Title  
  Series Editor Series Title Abbreviated Series Title  
  Series Volume Series Issue Edition  
  ISSN 1867-0334 ISBN Medium  
  Area Expedition Conference  
  Notes PMID:26514820 Approved no  
  Call Number NCSU @ edshirle @ Serial 3223  
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