کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
3369887 1219057 2007 4 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Automated extraction of avian influenza virus for rapid detection using real-time RT-PCR
موضوعات مرتبط
علوم زیستی و بیوفناوری ایمنی شناسی و میکروب شناسی میکروبیولوژی و بیوتکنولوژی کاربردی
پیش نمایش صفحه اول مقاله
Automated extraction of avian influenza virus for rapid detection using real-time RT-PCR
چکیده انگلیسی

BackgroundHighly pathogenic H5N1 avian influenza (AI) poses a grave risk to human health. An important aspect of influenza control is rapid diagnosis.ObjectivesThis 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 designAnalytical 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.ResultsBoth manual and robotic extractions detected AI virus at 1 log10 EID50/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.ConclusionThe 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.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Clinical Virology - Volume 40, Issue 2, October 2007, Pages 142–145
نویسندگان
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