Development and evaluation of tailored specific real-time RT-PCR assays for detection of foot-and-mouth disease virus serotypes circulating in East Africa
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Rapid, reliable and accurate diagnostic methods provide essential support to programmes that monitor and control foot-and-mouth disease (FMD). While pan-specific molecular tests for FMD virus (FMDV) detection are well established and widely used in endemic and FMD-free countries, current serotyping methods mainly rely either on antigen detection ELISAs or nucleotide sequencing approaches. This report describes the development of a panel of serotype-specific real-time RT-PCR assays (rRT-PCR) tailored to detect FMDV lineages currently circulating in East Africa. These assays target sequences within the VP1-coding region that share high intra-lineage identity, but do not cross-react with FMD viruses from other serotypes that circulate in the region. These serotype-specific assays operate with the same thermal profile as the pan-diagnostic tests making it possible to run them in parallel to produce CT values comparable to the pan-diagnostic test detecting the 3D-coding region. These assays were evaluated alongside the established pan-specific molecular test using field samples and virus isolates collected from Tanzania, Kenya and Ethiopia that had been previously characterised by nucleotide sequencing. Samples (n=71) representing serotype A (topotype AFRICA, lineage G-I), serotype O (topotypes EA-2 and EA-4), serotype SAT 1 (topotype I (NWZ)) and serotype SAT2 (topotype IV) were correctly identified with these rRT-PCR assays. Furthermore, FMDV RNA from samples that did not contain infectious virus could still be serotyped using these assays. These serotype-specific real-time RT-PCR assays can detect and characterise FMDVs currently circulating in East Africa and hence improve disease control in this region.
Original language | English |
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Journal | Journal of Virological Methods |
Volume | 237 |
Pages (from-to) | 114-120 |
Number of pages | 7 |
ISSN | 0166-0934 |
DOIs | |
Publication status | Published - Nov 2016 |
Externally published | Yes |
Bibliographical note
Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.
- Africa, Eastern/epidemiology, Animals, Capsid Proteins/genetics, Ethiopia/epidemiology, Foot-and-Mouth Disease/diagnosis, Foot-and-Mouth Disease Virus/classification, Kenya/epidemiology, Molecular Diagnostic Techniques/methods, RNA, Viral, Real-Time Polymerase Chain Reaction/methods, Sequence Analysis, DNA, Serogroup, Serotyping/methods, Tanzania/epidemiology
Research areas
ID: 257915103