An optimized method to detect influenza virus and human rhinovirus from exhaled breath and the airborne environment.
Respiratory viruses are difficult to characterize in the airborne environment due to their low concentration and the presence of a wide range of inhibitors. As a first step in studying airborne viruses, we optimized molecular biology methods to quantify influenza viruses and human rhinovirus. Quantitative PCR was used as an endpoint to evaluate RNA extraction techniques and reverse transcription protocols. We found that a Trizol-chloroform extraction and MultiScribe RT increased virus detection 10-fold compared to methods used in published field studies of airborne respiratory viruses. Virus was recovered without inhibition from samples contaminated with up to 50 microg/sample of particulate matter. The methods developed can be used in studies of airborne respiratory viruses.
Fabian
Patricia
P
Department of Environmental Health, Harvard School of Public Health, 401 Park Drive, Boston, MA, USA.
McDevitt
James Joseph
JJ
Lee
Wai-Ming
WM
Houseman
Eugene Andres
EA
Milton
Donald Kirby
DK
D000391:Air Microbiology; D005591:Chemical Fractionation / Q000379:methods; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000302:isolation & purification*; D052638:Particulate Matter / Q000737:chemistry; D012367:RNA, Viral / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012229:Rhinovirus / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D052638:Particulate Matter; D012367:RNA, Viral