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

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• 作者：Olamide T. Afolayan ; Cathy C. Webb ; Jennifer L. Cannon
• 关键词：Norovirus ; Porcine gastric mucin ; Ethanol ; Levulinic acid ; Sodium dodecyl sulfate ; Thermal inactivation ; Surrogate
• 刊名：Food and Environmental Virology
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：8
• 期：1
• 页码：70-78
• 全文大小：529 KB
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• 作者单位：Olamide T. Afolayan (1)
  Cathy C. Webb (1)
  Jennifer L. Cannon (1)
  
  1. Department of Food Science and Technology, Center for Food Safety, University of Georgia, 1109 Experiment Street, Griffin, GA, 30223, USA
  
• 刊物主题：Virology; Food Science; Chemistry/Food Science, general;
• 出版者：Springer US
• ISSN：1867-0342

文摘

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 °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 µg/ml PGM followed by RNase A (5 ng/µl) 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. Keywords Norovirus Porcine gastric mucin Ethanol Levulinic acid Sodium dodecyl sulfate Thermal inactivation Surrogate