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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
medicine
Development and evaluation of a real-time reverse transcription-loop- mediated isothermal amplification assay for rapid detection of rift valley fever virus in clinical specimens
Journal of Clinical Microbiology, Volume 47, No. 3, Year 2009
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Description
This paper reports on the development and validation of a real-time reverse transcription-loop-mediated isothermal amplification assay (RT-LAMP) targeting the genomic large RNA segment of Rift Valley fever virus (RVFV). The set of six designed RT-LAMP primers identified strains of RVFV isolated in geographically distinct areas over a period of 50 years; there was no cross-reactivity with other genetically related and unrelated arboviruses. When testing serial sera and plasma from sheep experimentally infected with wild-type RVFV, there was 100% agreement between results of the RT-LAMP, a TaqMan-based real-time PCR, and virus isolation. Similarly, the assay had very high levels of diagnostic sensitivity and specificity when testing various clinical specimens from humans and animals naturally infected with the virus during recent outbreaks of the disease in Africa. The detection of specific viral genome targets in positive clinical specimens was achieved in less than 30 min. As a highly accurate, rapid, and very simple nucleic acid detection format, the RT-LAMP has the potential to be used in less-well-equipped laboratories in Africa and as a portable device during RVF outbreaks in remote areas, and it can be a valuable tool for the differential diagnosis of viral hemorrhagic fevers. Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Authors & Co-Authors
le Roux, C.
South Africa, Johannesburg
National Institute for Communicable Diseases
South Africa, Pretoria
University of Pretoria
Kubo, T.
Japan, Nagasaki
Nagasaki University
Grobbelaar, Antoinette A.
South Africa, Johannesburg
National Institute for Communicable Diseases
Jansen Van Vuren, Petrus
South Africa, Johannesburg
National Institute for Communicable Diseases
Weyer, Jacqueline
South Africa, Johannesburg
National Institute for Communicable Diseases
Nel, Louis Hendrik
South Africa, Pretoria
University of Pretoria
Swanepoel, Robert
South Africa, Johannesburg
National Institute for Communicable Diseases
Morita, Kouichi
Japan, Nagasaki
Nagasaki University
Pawȩska, Janusz Tadeusz
South Africa, Johannesburg
National Institute for Communicable Diseases
South Africa, Johannesburg
School of Pathology
Statistics
Citations: 106
Authors: 9
Affiliations: 4
Identifiers
Doi:
10.1128/JCM.01412-08
ISSN:
00951137
Research Areas
Infectious Diseases