Propidium Monoazide Coupled with PCR Predicts Infectivity of Enteric Viruses in Swine Manure and Biofertilized Soil

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• 作者：Gislaine Fongaro ; Marta Hernández…
• 关键词：Infectious enteric viruses ; Recycle of swine manure ; Biofertilizer ; PMA–PCR
• 刊名：Food and Environmental Virology
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：8
• 期：1
• 页码：79-85
• 全文大小：435 KB
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• 作者单位：Gislaine Fongaro (1)
  Marta Hernández (2) (3)
  María Cruz García-González (2)
  Célia Regina Monte Barardi (1)
  David Rodríguez-Lázaro (4)
  
  1. Laboratório de Virologia Aplicada, Departamento de Microbiologia, Imunologia e Parasitologia, UFSC, Florianópolis, Brazil
  2. Instituto Tecnológico Agrario de Castilla y León, Valladolid, Spain
  3. Departamento de Ingeniería Agrícola y Forestal, Tecnología de los Alimentos, E.T.S. Ingenierías Agrarias, Universidad de Valladolid, 34004, Palencia, Spain
  4. Microbiology Section, Department of Biotechnology and Food Science, Universidad de Burgos, Burgos, Spain
  
• 刊物主题：Virology; Food Science; Chemistry/Food Science, general;
• 出版者：Springer US
• ISSN：1867-0342

文摘

The use of propidium monoazide (PMA) coupled with real-time PCR (RT-qPCR or qPCR for RNA or DNA viruses, respectively) was assessed to discriminate infectious enteric viruses in swine raw manure, swine effluent from anaerobic biodigester (AB) and biofertilized soils. Those samples were spiked either with infectious and heat-inactivated human adenovirus-2 (HAdV-2) or mengovirus (vMC₀), and PMA-qPCR/RT-qPCR allowed discriminating inactivated viruses from the infective particles, with significant reductions (>99.9 %). Then, the procedure was further assayed to evaluate the presence and stability of two non-cultivable viruses (porcine adenovirus and rotavirus A) in natural samples (swine raw manure, swine effluent from AB and biofertilized soils); it demonstrated viral inactivation during the storage period at 23 °C. As a result, the combination of PMA coupled to real-time PCR can be a promising alternative for prediction of viral infectivity in comparison to more labour-intensive and costly techniques such as animal or tissue-culture infectivity methods, and for those viruses that do not have currently available cell culture techniques. Keywords Infectious enteric viruses Recycle of swine manure Biofertilizer PMA–PCR