BACKGROUND: Highly pathogenic H5N1 avian influenza (AI) poses a grave risk to human health. An important aspect of influenza control is rapid diagnosis. OBJECTIVES: This study describes the efficiency of AI-RNA extraction utilizing silica-based magnetic beads with robotics and its detection with an influenza A matrix gene real-time RT-PCR from tracheal swabs, and compares it to virus isolation and manual spin column extractions. STUDY DESIGN: Analytical sensitivity was assessed by performing dilution analysis and detection of H2N2 AI viral RNA. Diagnostic sensitivity and specificity was assessed by analyzing tracheal swabs collected from H7N2 infected and uninfected chickens. RESULTS: Both manual and robotic extractions detected AI virus at 1log(10)EID(50)/ml. Diagnostic sensitivity and specificity of matrix gene detection with the automated extraction method for chicken tracheal swab specimens was similar to that of virus isolation and the manual extraction method. There were only three discordant results among 212 tested specimens. CONCLUSION: The main advantages of automated robotic viral nucleic acid extraction are high throughput processing; hands-free operation; and reduction in human and technical error. This study demonstrates successful detection of influenza A virus with magnetic beads utilizing the Qiagen MagAttract cell kit on a BioRobot M48 platform.
BACKGROUND: Highly pathogenic H5N1 avian influenza (AI) poses a grave risk to human health. An important aspect of influenza control is rapid diagnosis. OBJECTIVES: This study describes the efficiency of AI-RNA extraction utilizing silica-based magnetic beads with robotics and its detection with an influenza A matrix gene real-time RT-PCR from tracheal swabs, and compares it to virus isolation and manual spin column extractions. STUDY DESIGN: Analytical sensitivity was assessed by performing dilution analysis and detection of H2N2 AI viral RNA. Diagnostic sensitivity and specificity was assessed by analyzing tracheal swabs collected from H7N2 infected and uninfected chickens. RESULTS: Both manual and robotic extractions detected AI virus at 1log(10)EID(50)/ml. Diagnostic sensitivity and specificity of matrix gene detection with the automated extraction method for chicken tracheal swab specimens was similar to that of virus isolation and the manual extraction method. There were only three discordant results among 212 tested specimens. CONCLUSION: The main advantages of automated robotic viral nucleic acid extraction are high throughput processing; hands-free operation; and reduction in human and technical error. This study demonstrates successful detection of influenza A virus with magnetic beads utilizing the Qiagen MagAttract cell kit on a BioRobot M48 platform.
Authors: Darrell P Chandler; Sara B Griesemer; Christopher G Cooney; Rebecca Holmberg; Nitu Thakore; Becca Mokhiber; Phillip Belgrader; Christopher Knickerbocker; Jeanmarie Schied; Kirsten St George Journal: J Virol Methods Date: 2012-03-07 Impact factor: 2.014
Authors: Nicholas M Adams; Amy E Creecy; Catherine E Majors; Bathsheba A Wariso; Philip A Short; David W Wright; Frederick R Haselton Journal: Biomicrofluidics Date: 2013-01-18 Impact factor: 2.800
Authors: Jie He; Andrea J Kraft; Jiang Fan; Meredith Van Dyke; Lihua Wang; Michael E Bose; Marilyn Khanna; Jacob A Metallo; Kelly J Henrickson Journal: Viruses Date: 2009 Impact factor: 5.048