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標題: Development of a loop-mediated isothermal amplification for rapid detection of orf virus
作者: Tsai, S.M.
Chan, K.W.
Hsu, W.L.
Chang, T.J.
Wong, M.L.
Wang, C.Y.
關鍵字: orf virus;Loop-mediated isothermal amplification;(LAMP);PCR;Nested;PCR;polymerase-chain-reaction;reverse-transcription;mouth-disease;parapoxvirus;diagnosis;infection;goats
Project: Journal of Virological Methods
期刊/報告no:: Journal of Virological Methods, Volume 157, Issue 2, Page(s) 200-204.
A loop-mediated isothermal amplification (LAMP) assay using six primers targeting a highly conserved region of the B2L gene has been developed to diagnose orf virus. The assay produces a ladder-like pattern of products on an agarose gel that can be specifically digested with BsrGI enzyme. The sensitivity of the LAMP assay, which was determined to be a single copy of the standard plasmid, was 100 fold and 10 fold higher than PCR and nested PCR, respectively; furthermore, no cross-reactivity was founded with the other tested viruses. By staining the products directly in the tube with PicoGreen or ethidium bromide, the products can be visualized with a similar sensitivity as by gel electrophoresis. Clinical samples were tested using PCR, nested PCR and LAMP assay and the positive rates were 60%, 70% and 70%, respectively. The LAMP assay allows easy, rapid, accurate and sensitive detection of infection with orf virus and is especially applicable in a resource-limited situation. (C) 2009 Elsevier B.V. All rights reserved.
ISSN: 0166-0934
DOI: 10.1016/j.jviromet.2009.01.003
Appears in Collections:微生物暨公共衛生學研究所

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