Research Article Detection of Influenza Virus Infection Using Two PCR Methods Richard K. Zimmerman, 1 Charles R. Rinaldo, 2,3 Mary Patricia Nowalk, 1 G. K. Balasubramani, 4 Mark G. Thompson, 5 Arlene Bullotta, 2 Michael Susick, 1 and Stephen Wisniewski 4 1 Department of Family Medicine, University of Pittsburgh School of Medicine, 3518 5th Avenue, Pittsburgh, PA 15261, USA 2 Department of Pathology, University of Pittsburgh School of Medicine, PA 15261, USA 3 Department of Infectious Disease and Microbiology, University of Pittsburgh Graduate School of Public Health, Pittsburgh, PA 15261, USA 4 Department of Epidemiology, University of Pittsburgh Graduate School of Public Health, Pittsburgh, PA 15261, USA 5 Infuenza Division, Centers for Disease Control and Prevention, Atlanta, GA 30307, USA Correspondence should be addressed to Mary Patricia Nowalk; tnowalk@pitt.edu Received 15 September 2014; Accepted 21 November 2014; Published 9 December 2014 Academic Editor: Stefan P¨ ohlmann Copyright © 2014 Richard K. Zimmerman et al. Tis is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Rapid, accurate, and cost-efective methods to identify the cause of respiratory tract infections are needed to maximize clinical beneft. Outpatients with acute respiratory illness were tested for infuenza using a singleplex reverse transcriptase polymerase chain reaction (SRT-PCR) method. A multiplex RT-PCR (MRT-PCR) method tested for infuenza and 17 other viruses and was compared with SRT-PCR using chi-square tests. Among 935 patients, 335 (36%) tested positive for infuenza A and infuenza B using SRT-PCR. Using MRT-PCR, 320 (34.2%) tested positive for infuenza A and infuenza B. Tis study supports MRT-PCR as a comparable method for detecting infuenza among patients seeking outpatient care for acute respiratory illnesses. 1. Background Each year millions of people are aficted with infuenza- associated respiratory tract infections, some of which lead patients to seek outpatient medical attention, referred to as medically attended acute respiratory infection (ARI). Rapid, accurate, and cost-efective methods are needed to maxi- mize clinical beneft of antivirals and reduce inappropriate antibiotic usage. New assay methods using multiplex reverse transcriptase polymerase chain reaction (MRT-PCR) are available that allow for relatively rapid detection of multiple virus types including infuenza [1, 2]. 2. Objective During the 2012-2013 infuenza season, the Centers for Dis- ease Control and Prevention (CDC)-funded, multicenter US Infuenza Vaccine Efectiveness (Flu VE) Network conducted a study designed to determine the efectiveness of the sea- son’s infuenza vaccine. Tat study used singleplex RT-PCR (SRT-PCR) to detect infuenza virus. Te University of Pitts- burgh site of the Flu VE Network also used MRT-PCR. Te purpose of this study was to compare the agreement between SRT-PCR and MRT-PCR for infuenza virus detection. 3. Study Design Te US Flu VE Network evaluated the efectiveness of the season’s infuenza vaccine using a test-negative, case-control study design [3, 4], where the proportion vaccinated among those who test positive for infuenza was compared with the proportion vaccinated among those who test negative. Participants in the current study were enrollees in the University of Pittsburgh site of the Flu VE study, described previously [5]. Eligibility criteria included patients aged ≥6 months as of 9/1/2012, seeking outpatient medical care for an upper respiratory illness of ≤7 days’ duration with cough, and not taking an infuenza antiviral before the visit. Tis prospective study was approved by the University of Pittsburgh Institutional Review Board. Hindawi Publishing Corporation Advances in Virology Volume 2014, Article ID 274679, 3 pages http://dx.doi.org/10.1155/2014/274679