Deposition of respiratory virus pathogens on frequently touched surfaces at airports

Ikonen, Niina; Savolainen-Kopra, Carita; Enstone, Joanne E.; Kulmala, Ilpo; Pasanen, Pertti; Salmela, Anniina; Salo, Satu; Nguyen-Van-Tam, Jonathan S.; Ruutu, Petri

Viral nucleic acid was extracted from 100 μl samples with the Qiagen Qiacube® instrument using RNeasy Mini Kit® (Qiagen, Hilden, Germany) following the manufacturer's instructions and was eluted in 50 μl. Random hexamer primers and RevertAid H Minus Reverse Transcriptase (Thermo Fisher Scientific, Massachusetts, USA) were used in the synthesis of the cDNA. cDNA reaction was performed at the following conditions: 10 min at 25°C, 30 min at 42°C and 10 min at 70°C. All samples were tested in three separate multiplex real-time polymerase chain reaction (real-time PCR) tests using QuantiTect™ Multiplex PCR or NoRox PCR Kit (Qiagen, Hilden, Germany). Primers and probes for seasonal influenza A [25] [26] [27] (with influenza A(H3)primer and probe sequences courtesy of Erasmus Medical Centel, Rotterdam, Netherlands) and B viruses [28] , respiratory syncytial virus [28] , adenovirus [29] , rhinovirus [30] and coronavirus (229E, HKU1, NL63 and OC43)