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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
immunology and microbiology
Development and evaluation of loop-mediated isothermal amplification assay for detection of Crimean Congo hemorrhagic fever virus in Sudan
Journal of Virological Methods, Volume 190, No. 1-2, Year 2013
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Description
Crimean-Congo hemorrhagic fever (CCHF) virus (CCHFV) activity has been detected in Kordufan region of the Sudan in 2008 with high case-fatality rates in villages and rural hospitals in the region. Therefore, in the present study, a reverse transcription (RT) loop-mediated isothermal amplification (RT-LAMP) assay was developed and compared to nested RT-PCR for rapid detection of CCHFV targeting the small (S) RNA segment. A set of RT-LAMP primers, designed from a highly conserved region of the S segment of the viral genome, was employed to identify all the Sudanese CCHFV strains. The sensitivity studies indicated that the RT-LAMP detected 10 fg of CCHFV RNA as determined by naked eye turbidity read out, which is more likely the way it would be read in a resource-poor setting. This level of sensitivity is good enough to detect most acute cases. Using agarose gel electrophoresis, the RT-LAMP assay detected as little as 0.1 fg of viral RNA (equivalent to 50 viral particle). There was 100% agreement between results of the RT-LAMP and the nested PCR when testing 10-fold serial dilution of CCHFV RNA. The specificity studies indicated that there was no cross-reactivity with other related hemorrhagic fever viruses circulating in Sudan including, Rift Valley fever virus (RVFV), Dengue fever virus, and yellow fever virus. The RT-LAMP was performed under isothermal conditions at 63 °C and no special apparatus was needed, which rendered the assay more economical and practical than real-time PCR in such developing countries, like Sudan. In addition, the RT-LAMP provides a valuable tool for rapid detection and differentiation of CCHFV during an outbreak of the disease in remote areas and in rural hospitals with resource-poor settings. © 2013 Elsevier B.V.
Authors & Co-Authors
Osman, Hana A.M.
Sudan, Khartoum
University of Khartoum Faculty of Veterinary Medicine
Sudan, Khartoum
Khartoum University
Eltom, Kamal Hassan
Sudan, Khartoum
University of Khartoum Faculty of Veterinary Medicine
Musa, Nasreen O.
Sudan, Khartoum
University of Khartoum Faculty of Veterinary Medicine
Bilal, Nasreldin M.
Sudan, Khartoum
Khartoum University
Elbashir, Mustafa Idris
Sudan, Khartoum
University of Khartoum Faculty of Medicine
Aradaib, Imadeldin Elamin
Sudan, Khartoum
University of Khartoum Faculty of Veterinary Medicine
Statistics
Citations: 31
Authors: 6
Affiliations: 3
Identifiers
Doi:
10.1016/j.jviromet.2013.03.004
ISSN:
01660934
e-ISSN:
18790984
Research Areas
Health System And Policy
Infectious Diseases
Study Locations
Congo
Sudan