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Detection and identification of Rift Valley fever virus in mosquito vectors by quantitative real-time PCR

Diagnostic methods allowing for rapid identification of pathogens are crucial for controlling and preventing dissemination after disease outbreaks as well as for use in surveillance programs. For arboviruses, detection of the presence of virus in their arthropod hosts is important for monitoring of...

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Main Authors: Mwaengo, D., Lorenzo, G., Iglesias, J., Warigia, M., Sang, R., Bishop, Richard P., Brun, A.
Format: Journal Article
Language:Inglés
Published: Elsevier 2012
Subjects:
animal diseases
parasites
livestock
Online Access:https://hdl.handle.net/10568/21585
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author Mwaengo, D.
Lorenzo, G.
Iglesias, J.
Warigia, M.
Sang, R.
Bishop, Richard P.
Brun, A.
author_browse Bishop, Richard P.
Brun, A.
Iglesias, J.
Lorenzo, G.
Mwaengo, D.
Sang, R.
Warigia, M.
author_facet Mwaengo, D.
Lorenzo, G.
Iglesias, J.
Warigia, M.
Sang, R.
Bishop, Richard P.
Brun, A.
author_sort Mwaengo, D.
collection Repository of Agricultural Research Outputs (CGSpace)
description Diagnostic methods allowing for rapid identification of pathogens are crucial for controlling and preventing dissemination after disease outbreaks as well as for use in surveillance programs. For arboviruses, detection of the presence of virus in their arthropod hosts is important for monitoring of viral activity and quantitative information is useful for modeling of transmission dynamics. In this study, molecular detection of Rift Valley fever virus (RVFV) in mosquito samples from the 2006 to 2007 East African outbreaks was performed using quantitative real-time PCR assay (qRT-PCR). Specific RVFV sequence-based primer/fluorogenic (TaqMan) probe sets were derived from the L and S RNA segments of the virus. Both primer-probe L and S segment-based combinations detected genomic RVFV sequences, with generally comparable levels of sensitivity. Viral loads from three mosquito species, Aedes mcintoshi, Aedes ochraceus and Mansonia uniformis were estimated and significant differences of between 5- and 1000-fold were detected between Ae. mcintoshi and M. uniformis using both the L and S primer-probe-based assays. The genetic relationships of the viral sequences in mosquito samples were established by partial M segment sequencing and assigned to the two previously described viral lineages defined by analysis of livestock isolates obtained during the 2006–2007 outbreak, confirming that similar viruses were present in both the vector and mammalian host. The data confirms the utility of qRT-PCR for identification and initial quantification of virus in mosquito samples during RVFV outbreaks.
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language Inglés
publishDate 2012
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spelling CGSpace215852025-06-13T04:20:20Z Detection and identification of Rift Valley fever virus in mosquito vectors by quantitative real-time PCR Mwaengo, D. Lorenzo, G. Iglesias, J. Warigia, M. Sang, R. Bishop, Richard P. Brun, A. animal diseases parasites livestock Diagnostic methods allowing for rapid identification of pathogens are crucial for controlling and preventing dissemination after disease outbreaks as well as for use in surveillance programs. For arboviruses, detection of the presence of virus in their arthropod hosts is important for monitoring of viral activity and quantitative information is useful for modeling of transmission dynamics. In this study, molecular detection of Rift Valley fever virus (RVFV) in mosquito samples from the 2006 to 2007 East African outbreaks was performed using quantitative real-time PCR assay (qRT-PCR). Specific RVFV sequence-based primer/fluorogenic (TaqMan) probe sets were derived from the L and S RNA segments of the virus. Both primer-probe L and S segment-based combinations detected genomic RVFV sequences, with generally comparable levels of sensitivity. Viral loads from three mosquito species, Aedes mcintoshi, Aedes ochraceus and Mansonia uniformis were estimated and significant differences of between 5- and 1000-fold were detected between Ae. mcintoshi and M. uniformis using both the L and S primer-probe-based assays. The genetic relationships of the viral sequences in mosquito samples were established by partial M segment sequencing and assigned to the two previously described viral lineages defined by analysis of livestock isolates obtained during the 2006–2007 outbreak, confirming that similar viruses were present in both the vector and mammalian host. The data confirms the utility of qRT-PCR for identification and initial quantification of virus in mosquito samples during RVFV outbreaks. 2012-10 2012-08-16T08:52:21Z 2012-08-16T08:52:21Z Journal Article https://hdl.handle.net/10568/21585 en Limited Access Elsevier Mwaengo, D., Lorenzo, G., Iglesias, J., Warigia, M., Sang, R., Bishop, R.P. and Brun, A. 2012. Detection and identification of Rift Valley fever virus in mosquito vectors by quantitative real-time PCR. Virus Research 169(1): 137-143.
spellingShingle animal diseases
parasites
livestock
Mwaengo, D.
Lorenzo, G.
Iglesias, J.
Warigia, M.
Sang, R.
Bishop, Richard P.
Brun, A.
Detection and identification of Rift Valley fever virus in mosquito vectors by quantitative real-time PCR
title Detection and identification of Rift Valley fever virus in mosquito vectors by quantitative real-time PCR
title_full Detection and identification of Rift Valley fever virus in mosquito vectors by quantitative real-time PCR
title_fullStr Detection and identification of Rift Valley fever virus in mosquito vectors by quantitative real-time PCR
title_full_unstemmed Detection and identification of Rift Valley fever virus in mosquito vectors by quantitative real-time PCR
title_short Detection and identification of Rift Valley fever virus in mosquito vectors by quantitative real-time PCR
title_sort detection and identification of rift valley fever virus in mosquito vectors by quantitative real time pcr
topic animal diseases
parasites
livestock
url https://hdl.handle.net/10568/21585
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