Proceedings

Rapid, sensitive and effective diagnostic tools for foot-and-mouth disease virus in Africa

Christopher J. Kasanga, Wataru Yamazaki, Valerie Mioulet, Donald P. King, Misheck Mulumba, Ezekia Ranga, Jimis Deve, Cornelius Mundia, Patrick Chikungwa, Laureta Joao, Philemon N. Wambura, Mark M. Rweyemamu
Onderstepoort Journal of Veterinary Research | Vol 81, No 2 | a727 | DOI: https://doi.org/10.4102/ojvr.v81i2.727 | © 2014 Christopher J. Kasanga, Wataru Yamazaki, Valerie Mioulet, Donald P. King, Misheck Mulumba, Ezekia Ranga, Jimis Deve, Cornelius Mundia, Patrick Chikungwa, Laureta Joao, Philemon N. Wambura, Mark M. Rweyemamu | This work is licensed under CC Attribution 4.0
Submitted: 05 December 2013 | Published: 23 April 2014

About the author(s)

Christopher J. Kasanga, Southern African Centre for Infectious Disease Surveillance, Sokoine University of Agriculture, Tanzania, United Republic of
Wataru Yamazaki, Department of Microbiology, University of Miyazaki, Japan
Valerie Mioulet, The Pirbright Institute, United Kingdom
Donald P. King, The Pirbright Institute, United Kingdom
Misheck Mulumba, Southern African Development Community Secretariat, Private Bag 0095, Gaborone, Botswana, Botswana
Ezekia Ranga, Ministry of Livestock Development and Fisheries, Tanzania, United Republic of
Jimis Deve, Southern African Development Community, Transboundary Animal Diseases Section, Tanzania, United Republic of
Cornelius Mundia, Southern African Development Community, Transboundary Animal Diseases Section, Zambia
Patrick Chikungwa, Southern African Development Community, Transboundary Animal Diseases Section, Malawi
Laureta Joao, Southern African Development Community, Transboundary Animal Diseases Section, Angola
Philemon N. Wambura, Ministry of Livestock Development and Fisheries, Tanzania, United Republic of
Mark M. Rweyemamu, Southern African Centre for Infectious Disease Surveillance, Sokoine University of Agriculture, Tanzania, United Republic of

Abstract

Speed is paramount in the diagnosis of highly infectious diseases, such as foot-and-mouth disease (FMD), as well as for emerging diseases; however, simplicity is required if a test is to be deployed in the field. Recent developments in molecular biology have enabled the specific detection of FMD virus (FMDV) by reverse-transcription loop-mediated isothermal amplification (RT-LAMP), real-time  reverse-transcription polymerase chain reaction (RT-qPCR) and sequencing. RT-LAMP enables amplification of the FMDV RNA-dependent RNA polymerase 3D(pol) gene at 63 °C (in the presence of a primer mixture and both reverse transcriptase and Bst DNA polymerase) for 1 h, whilst RT-qPCR amplifies the same gene in approximately 2 h 30 min. In this study, we compared the sensitivity and effectiveness of RT-LAMP against RT-qPCR for the detection of the FMDV 3D(pol) gene in 179 oesophageal-pharyngeal scraping samples (collected by probang) obtained from clinically healthy cattle and buffalo in Malawi, Mozambique and Tanzania in 2010. The FMDV detection rate was higher with RT-LAMP (30.2%; n = 54) than with RT-qPCR (17.3%; n = 31). All samples positive by RT-qPCR (Cq ≤ 32.0) were also positive for the RT-LAMP assay; and both assays proved to be highly specific for the FMDV target sequence. In addition, the VP1 sequences of 10 viruses isolated from positive samples corresponded to the respective FMDV serotypes and genotypes. Our findings indicate that the performance of RT-LAMP is superior to RT-qPCR. Accordingly, we consider this test to have great potential with regard to the specific detection and surveillance of infectious diseases of humans and animals in resource-compromised developing countries.

Keywords

Foot-and-mouth disease virus; RT-qPCR; RT-LAMP; diagnostics; Southern Africa

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