, 2011). Participants in the fitted N95 arm underwent a fit testing procedure using a 3M™ ABT 199 FT-30 Bitrex Fit Test Kit according to the manufacturers’

instructions (3M™, St Paul, MN, USA) (MacIntyre et al., 2011). All participants were followed up for four weeks for development of respiratory symptoms, and for an additional week after mask wearing had ceased (to account for incubation of infections acquired in week 4). Validated diary cards were provided for the four-week period to record daily the (1) number of hours worked; (2) mask/respirator usage; and (3) recognized CRI (MacIntyre et al., 2011). Participants were contacted daily by the study team either by phone or face-to-face contact to actively identify incident cases of viral respiratory infection. CRI was defined as at least two respiratory symptoms (cough, sneezing, runny nose, Anti-diabetic Compound Library ic50 shortness of breath, sore throat) or one respiratory symptom and one systemic symptom (including fever, headache, and lethargy). If any respiratory symptom was present, subjects were tested, following collection of a nose and throat swab, for bacterial and viral pathogens. Subjects with respiratory symptoms had two pharyngeal swabs collected by a trained nurse or doctor. Double rayon-tipped, plastic-shafted swabs were used to scratch both tonsil areas and the posterior

pharyngeal wall. These were transported immediately after collection to the laboratory, or at 4 °C if transport was delayed within 48 h. Pharyngeal swabs were tested at the Laboratories of the Beijing Centers for Disease

Control and Prevention. A multiplex PCR (Seegen Inc., Seoul, Korea) was used to detect S. pneumoniae, M. pneumoniae, B. pertussis, Legionella spp., Chlamydia and H. influenza type B. After aminophylline preheating at 95 °C for 15 min, 40 amplification cycles were carried out under the following conditions in a thermal cycler (GeneAmp PCR system 9700, Foster City, CA, USA): 94 °C for 30 s, 60 °C for 1.5 min, and 72 °C for 1.5 min. Amplification was completed at the final extension step at 72 °C for 10 min. The multiplex PCR products were visualized by electrophoresis on an ethidium bromide-stained 2% agarose gel. Laboratory-confirmed viral respiratory infection, defined as detection of adenoviruses, human metapneumovirus, coronaviruses 229E/NL63 and OC43/HKU1, parainfluenza viruses 1, 2 and 3, influenza viruses A and B, respiratory syncytial viruses A and B, or rhinovirus A/B by nucleic acid testing (NAT) using a commercial multiplex polymerase chain reaction (PCR) (Seegen, Inc., Seoul, Korea) as previously described ( MacIntyre et al., 2011). The endpoint of interest, bacterial colonization and co-infection with two bacteria or virus and bacteria were analyzed by intention-to-treat analysis.