期刊
MOLECULAR AND CELLULAR PROBES
卷 23, 期 2, 页码 71-74出版社
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.mcp.2008.12.001
关键词
Classical swine fever virus (CSFV); Reverse transcription of loop-mediated isothermal amplification (RT-LAMP); RT-PCR; Pre-clinical detection
类别
资金
- National Key Technologies R&D Program of China
- National Natural Science Foundation of China [30671563, 30700597]
The usefulness of reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid preclinical detection of classical swine fever virus (CSFV) infection was evaluated. The RT-LAMP reaction could be finished in 60 min under isothermal condition at 65 degrees C by employing a set of four primers targeting the 50 untranslated region of CSFV. The RT-LAMP assay of CSFV showed higher sensitivities than that of RT-PCR, with a detection limit of 5 copies per reaction. No cross-reactivity was observed from the samples of other related viruses including porcine circovirus type 2, porcine parvovirus, porcine pseudorabies virus, Japanese encephalitis virus, and porcine reproductive and respiratory syndrome virus. The detection rates of CSFV RT-LAMP, RT-PCR and virus isolation for samples including blood, tonsil, nasal and rectal swabs from uninoculated pigs without any clear clinical symptom were 89%, 78% and 71%, respectively. Furthermore, all of the assays showed higher sensitivity for blood and tonsil swabs samples than nasal and rectal swabs. These results indicate that the CSFV RT-LAMP assay is a valuable tool for its rapid, cost-effective detection and has potential usefulness for rapid pre-clinical detection and surveillance of classical swine fever in developing countries. (C) 2008 Elsevier Ltd. All rights reserved.
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