Protocol for Metagenomic Virus Detection in Clinical Specimens

Ethics Statement Production of specific pathogen–free eggs (VALO BioMedia GmbH, OsterholzScharmbeck, Germany) was performed in accordance with the guidelines of the European Pharmacopoeia (EP7.0.5.2.2.) and the US Department of Agriculture Veterinary Services (Memorandum 800.65). All procedures regarding embryonated chicken eggs were in accordance with the German Animal Protection Law. For infection, fertilized chicken eggs at embryonation day 11 were inoculated with virus into the allantois sack or onto the chorioallantoic membrane. Development of embryos was terminated at the 17th day of embryonation by cooling the eggs overnight at 4°C. No further specific approval is needed for experiments on embryonated birds before the time of hatching.