Rutjes Saskia A, Lodder-Verschoor Froukje, van der Poel Wim H M, van Duijnhoven Yvonne T H P, de Roda Husman Ana Maria
Microbiological Laboratory for Health Protection, Centre for Infectious Disease Control, National Institute of Public Health and the Environment, P.O. Box 1/ Antonie van Leeuwenhoeklaan 9, 3720 BA Bilthoven, The Netherlands.
J Food Prot. 2006 Aug;69(8):1949-56. doi: 10.4315/0362-028x-69.8.1949.
Disease outbreaks in which foods are epidemiologically implicated as the common source are frequently reported. Noroviruses and enteric hepatitis A viruses are among the most prevalent causative agents of foodborne diseases. However, the detection of these viruses in foods other than shellfish is often time-consuming and unsuccessful. In this study, three virus concentration methods were compared: polyethylene glycol (PEG) plus NaCl, ultracentrifugation, and ultrafiltration. Two RNA extraction methods, TRIzol and RNeasy Mini Kit (Qiagen), were compared for detection of viruses in whipped cream and lettuce (as representatives of the dairy and vegetable-fruit food groups, respectively). A seeding experiment with canine calicivirus was conducted to determine the efficiency of each virus extraction procedure. The PEG-NaCl-TRIzol method was most efficient for the detection of viruses in whipped cream and the ultracentrifugation-RNeasy-Mini Kit procedure was best for detection on lettuce. Based on the seeding experiments, food items implicated in norovirus-associated gastroenteritis outbreaks were subjected to the optimal procedure for a specific composition and matrix. No noroviruses were detected in the implicated food items, possibly because the concentration of virus on the food item was too low or because of the presence of inhibitory factors. For each food group, a specific procedure is optimal. Inhibitory factors should be controlled in these procedures because they influence virus detection in food.
经常有报道称,食物在疾病暴发中被认定为共同传染源。诺如病毒和甲型肝炎病毒是食源性疾病最常见的病原体。然而,在除贝类之外的食物中检测这些病毒往往既耗时又难以成功。在本研究中,比较了三种病毒浓缩方法:聚乙二醇(PEG)加氯化钠、超速离心和超滤。还比较了两种RNA提取方法,即TRIzol和RNeasy Mini试剂盒(Qiagen),分别用于检测生奶油和生菜(分别作为乳制品和蔬菜水果类食物的代表)中的病毒。进行了一项犬杯状病毒接种实验,以确定每种病毒提取方法的效率。PEG-氯化钠-TRIzol方法在检测生奶油中的病毒时效率最高,超速离心-RNeasy Mini试剂盒方法在检测生菜中的病毒时效果最佳。基于接种实验,将与诺如病毒相关的胃肠炎暴发中涉及的食品按照特定的成分和基质采用最佳程序进行处理。在所涉及的食品中未检测到诺如病毒,可能是因为食品上病毒的浓度过低,或者是因为存在抑制因子。对于每个食品类别,都有一个特定的最佳程序。在这些程序中应控制抑制因子,因为它们会影响食品中病毒的检测。