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標題: Development of a quantitative Light Cycler real-time RT-PCR for detection of avian reovirus
作者: Ke, G.M.
Cheng, H.L.
Ke, L.Y.
Ji, W.T.
Chulu, J.L.C.
Liao, M.H.
Chang, T.J.
Liu, H.J.
關鍵字: avian reovirus;Light Cyler;polymerase chain reaction;polymerase-chain-reaction;fragment length polymorphism;protein;sigma-ns;genome segment;phylogenetic analysis;binding activity;infected-cells;gene;identification;s1133
Project: Journal of Virological Methods
期刊/報告no:: Journal of Virological Methods, Volume 133, Issue 1, Page(s) 6-13.
A robust, ultrasensitive, and accurate quantitative assay was developed for avian reovirus (ARV) with the Light Cycler SYBR Green-based real-time reverse transcription-PCR (real-time LC RT-PCR). The assay exhibited high specificity as all negative controls and other avian pathogens, such as Newcastle disease virus (NDV), infectious bronchitis virus (IBV), infectious bursaL disease virus (IBDV), avian influenza virus (AIV), and mycoplasma synovia (NIS), failed to show any positive detection. A minimum of 39 copies/mu L of ARV genomic RNA could be detected by the assay. By dilution analysis, the real-time LC RT-PCR developed in this study was 3-log more sensitive than the conventional RT-PCR for the detection of ARV. The vaccine and field isolates of ARV were detected by the real-time LC RT-PCR. As a result of the high sensitivity and specificity of the assay with a relatively rapid and simple procedure, the real-time LC RT-PCR will be useful as a routine assay for the clinical diagnosis of ARV infection. (c) 2005 Elsevier B.V. All rights reserved.
ISSN: 0166-0934
DOI: 10.1016/j.jviromet.2005.09.011
Appears in Collections:分子生物學研究所

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