Foot-and-mouth disease (FMD), an economically important disease of cloven-hoofed animals, is endemic in Pakistan where three virus serotypes are present (O, A and Asia 1). Fifty-eight clinical samples collected between 2005 and 2008 from animals with suspected FMD in various locations in Pakistan were subjected to virus isolation on primary cell culture, antigen ELISA and real-time RT-PCR (rRT-PCR). Viruses were isolated from 32 of these samples and identified as FMDV type O (n = 31) or type A (n = 1). Foot-and-mouth disease virus (FMDV) genome was detected in a further 11 samples by real-time RT-PCR. Phylogenetic analyses of the VP1 nucleotide sequences showed that all of the type O viruses belonged to the MIDDLE EAST-SOUTH ASIA topotype with the majority belonging to the PanAsia-2 lineage; a single example of the older PanAsia lineage was identified. The single FMDV type A virus belonged to the ASIA topotype, but did not cluster with known strains that are currently circulating (such as Iran-05) and was not closely related to other type A viruses from the region. These findings demonstrate the widespread distribution of O-PanAsia-2 in Pakistan and the presence of undisclosed novel type A lineages in the region.

The aim of this study was to characterize foot-and-mouth disease (FMD) viruses collected between 2004 and 2008 from Sudan, a country where FMD is endemic. Using virus isolation and antigen ELISA, three FMD virus serotypes (O, A and SAT2) were detected in 24 samples that were submitted to the FAO World Reference Laboratory for FMD. Pan-serotypic real-time RT-PCR assays targeting the 5' untranslated region (5'UTR) and 3D genes of FMD virus were also used to contribute to the laboratory diagnosis of these cases. The lack of concordant results between the real-time RT-PCR assays for three serotype O viruses was attributed to four nucleotide mismatches in the 5'UTR PCR primer and probe sites (three substitutions for the sense-primer and one in the TaqMan(®) probe region). Taken together, the laboratory results showed that recent FMD outbreaks that occurred during 2008 in northern and central Sudan were caused by serotypes O and SAT2, while serotype A was last detected in 2006. Phylogenetic analyses of VP1 sequences from these viruses were used to determine the relationships with 23 older viruses from Sudan and other viruses from West and East Africa. For serotype O, closest genetic identities were between concurrent and historical Sudanese isolates, indicating that within-country circulation is an important mechanism by which FMD is maintained year-on-year in Sudan. A similar pattern was also evident for serotype A and SAT2 viruses; however, these lineages also contained recent representative FMD viral isolates from other countries in the region suggesting that long-distance animal movement can also contribute to FMD dispersal across sub-Saharan Africa. These findings provide the first molecular description of FMD viruses that are circulating in Sudan, and highlight that further sampling of representative viruses from the region is required before the complex epidemiology of FMD in sub-Saharan Africa can be fully understood.

A loop-mediated isothermal amplification (LAMP) assay was developed for the detection of African swine fever virus (ASFV). This assay targets the topoisomerase II gene of ASFV and its specificity was confirmed by restriction enzyme digestion of the reaction products. The analytical sensitivity of this ASFV LAMP assay was at least 330 genome copies, and the test was able to detect representative isolates of ASFV (n=38) without cross-reacting with classical swine fever virus. The performance of the LAMP assay was compared with other laboratory tests used for ASF diagnosis. Using blood and tissue samples collected from pigs experimentally infected with ASFV (Malawi isolate), there was good concordance between the LAMP assay and real-time PCR. In addition to detecting the reaction products using either agarose gels or real-time PCR machines, it was possible to visualise dual-labelled biotin and fluorescein ASFV LAMP amplicons using novel lateral flow devices. This assay and detection format represents the first step towards developing a practical, simple-to-use and inexpensive molecular assay format for ASF diagnosis in the field which is especially relevant to Africa where the disease is endemic in many countries.

In 2007, serological evidence for foot-and-mouth disease (FMD) infection was found as a result of differential diagnostic testing of Cypriot sheep suspected to be infected with bluetongue or contagious ecthyma. Seropositive sheep and goats were subsequently uncovered on ten geographically clustered flocks, while cattle and pigs in neighbouring herds were all seronegative. These antibodies were specific for serotype-O FMD virus, reacting with both structural and nonstructural (NS) FMD viral proteins. However, no FMD virus could be recovered from the seropositive flocks. FMD had not been recorded in Cyprus since 1964 and there has been no vaccination programme since 1984. Since all the seropositive animals were at least 3 years old and home-bred, it was concluded that infection had occurred approximately 3 years previously had passed unnoticed and died out spontaneously. It therefore appears that antibodies to FMD virus NS proteins can still be detected around 3 years after infection of small ruminants, but that virus carriers cannot be detected at this time. This unusual situation of finding evidence of historical infection in a FMD-free country caused considerable disruption and alarm and posed questions about the definition of what constitutes a FMD outbreak.

Nine viral diseases included in the World Organization for Animal Health list of notifiable diseases (former list A) were chosen for their contagiousness and high capacity of spreading to improve their diagnosis using new and emerging technologies. All the selected diseases - foot-and-mouth disease, swine vesicular disease, vesicular stomatitis, classical swine fever, African swine fever, bluetongue, African horse sickness, Newcastle disease and highly pathogenic avian influenza - are considered as transboundary diseases, which detection causes the prohibition of livestock exportation, and, thus, it leads to high economical losses. The applied diagnostic techniques can fall into two categories: (i) nucleic-acid detection, including padlock probes, real-time PCR with TaqMan, minor groove binding probes and fluorescence energy transfer reaction probes, isothermal amplification like the Cleavase/Invader assay or the loop-mediated amplification technology and the development of rapid kits for 'mobile' PCR and (ii) antigen-antibody detection systems like simplified and more sensitive ELISA tests. Besides, internal controls have been improved for nucleic acid-detecting methods by using an RNA plant virus - Cowpea Mosaic Virus - to ensure the stability of the RNA used as a positive control in diagnostic real-time RT-PCR assays. The development of these diagnosis techniques has required the joint efforts of a European consortium in which nine diagnostic laboratories and an SME who have collaborated since 2004 within the European Union-funded Lab-on-site project. The results obtained are shown in this paper.

Veterinary RecordVolume 162, Issue 10 p. 315-316 Short Communication Prospects for rapid diagnosis of foot-and-mouth disease in the field using reverse transcriptase-pcr D. P. King PhD, D. P. King PhD Institute for Animal Health, Ash Road, Pirbright, Surrey, GU24 0NFSearch for more papers by this authorJ. P. Dukes PhD, J. P. Dukes PhD Institute for Animal Health, Ash Road, Pirbright, Surrey, GU24 0NFSearch for more papers by this authorS. M. Reid PhD, S. M. Reid PhD Institute for Animal Health, Ash Road, Pirbright, Surrey, GU24 0NFSearch for more papers by this authorK. Ebert BSc, K. Ebert BSc Institute for Animal Health, Ash Road, Pirbright, Surrey, GU24 0NFSearch for more papers by this authorA. E. Shaw BSc, A. E. Shaw BSc Institute for Animal Health, Ash Road, Pirbright, Surrey, GU24 0NFSearch for more papers by this authorC. E. Mills PhD, C. E. Mills PhD Smiths Detection, 459 Park Avenue, Bushey, Watford, WD23 2BWSearch for more papers by this authorL. Boswell PhD, L. Boswell PhD Smiths Detection, 459 Park Avenue, Bushey, Watford, WD23 2BWSearch for more papers by this authorN. P. Ferris PhD, N. P. Ferris PhD Institute for Animal Health, Ash Road, Pirbright, Surrey, GU24 0NFSearch for more papers by this author D. P. King PhD, D. P. King PhD Institute for Animal Health, Ash Road, Pirbright, Surrey, GU24 0NFSearch for more papers by this authorJ. P. Dukes PhD, J. P. Dukes PhD Institute for Animal Health, Ash Road, Pirbright, Surrey, GU24 0NFSearch for more papers by this authorS. M. Reid PhD, S. M. Reid PhD Institute for Animal Health, Ash Road, Pirbright, Surrey, GU24 0NFSearch for more papers by this authorK. Ebert BSc, K. Ebert BSc Institute for Animal Health, Ash Road, Pirbright, Surrey, GU24 0NFSearch for more papers by this authorA. E. Shaw BSc, A. E. Shaw BSc Institute for Animal Health, Ash Road, Pirbright, Surrey, GU24 0NFSearch for more papers by this authorC. E. Mills PhD, C. E. Mills PhD Smiths Detection, 459 Park Avenue, Bushey, Watford, WD23 2BWSearch for more papers by this authorL. Boswell PhD, L. Boswell PhD Smiths Detection, 459 Park Avenue, Bushey, Watford, WD23 2BWSearch for more papers by this authorN. P. Ferris PhD, N. P. Ferris PhD Institute for Animal Health, Ash Road, Pirbright, Surrey, GU24 0NFSearch for more papers by this author First published: 08 March 2008 https://doi.org/10.1136/vr.162.10.315Citations: 11Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL No abstract is available for this article.Citing Literature Volume162, Issue10March 2008Pages 315-316 RelatedInformation