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Objectives Arboviruses pose a significant global health challenge. This study investigated the seroprevalence of major human arboviral infections, including yellow fever (YFV), dengue (DENV), Crimean-Congo hemorrhagic fever (CCHF), Rift Valley fever (RVF), West Nile virus (WNV), and chikungunya (CHIK), in Darfur region from September to December 2018. ELISA-IgM was used to detect antibodies. RT‒PCR was used to differentiate YFV infection from vaccine-immuno-response in IgM samples. Results A total of 152 blood samples were collected, with 123 (80.9%) from males and 29 (19.1%) from females. The participants were grouped by age: 50 (32.9%) were under 20 years, 96 (63.2%) were aged 20–45 years, and 6 (3.9%) were over 45 years. The seroprevalence rates for YFV, DENV, and CHIKV were 68 (44.7%), 23 (15.1%), and 5 (3.3%), respectively. There were 11 molecularly-confirmed YFV cases (7.2%). Among these, 3/11 were positive for DENV-IgM, and 1/11 was positive for CHIKV-IgM. Among the 68 YFV-positive individuals, 15 (22.1%) had been exposed to DENV, and 2 (2.9%) had been exposed to CHIKV. Co-exposure to DENV and CHIKV was detected in 3 (1.9%) patients, while 2 (1.3%) patients had triple exposure to YFV, CHIKV, or DENV. No exposure to CCHF, RVFV, or WNV was detected.

Q fever (QF) and Rift Valley fever (RVF) are endemic zoonotic diseases in African countries, causing significant health and economic burdens. Accurate prevalence estimates, crucial for disease control, rely on robust diagnostic tests. While enzyme-linked immunosorbent assays (ELISA) are not the gold standard, they offer rapid, cost-effective, and practical alternatives. However, varying results from different tests and laboratories can complicate comparing epidemiological studies. This study aimed to assess the agreement of test results for QF and RVF in humans and livestock across different laboratory conditions and, for humans, different types of diagnostic tests. We measured inter-laboratory agreement using concordance, Cohen’s kappa, and prevalence and bias-adjusted kappa (PABAK) on 91 human and 102 livestock samples collected from rural regions in Chad. The serum aliquots were tested using ELISA in Chad, and indirect immunofluorescence assay (IFA) (for human QF and RVF) and ELISA (for livestock QF and RVF) in Switzerland and Germany. Additionally, we examined demographic factors influencing test agreement, including district, setting (village vs. camp), sex, age, and livestock species of the sampled individuals. The inter-laboratory agreement ranged from fair to moderate. For humans, QF concordance was 62.5%, Cohen’s kappa was 0.31, RVF concordance was 81.1%, and Cohen’s kappa was 0.52. For livestock, QF concordance was 92.3%, Cohen’s kappa was 0.59, RVF concordance was 94.0%, and Cohen’s kappa was 0.59. Multivariable analysis revealed that QF test agreement is significantly higher in younger humans and people living in villages compared to camps and tends to be higher in livestock from Danamadji compared to Yao, and in small ruminants compared to cattle. Additionally, RVF agreement was found to be higher in younger humans. Our findings emphasize the need to consider sample conditions, test performance, and influencing factors when conducting and interpreting epidemiological seroprevalence studies.

In this research, we describe the first aptasensor for the detection of the Rift Valley Fever virus (RVFV). The process involved the selection of aptamers through the systematic evolution of ligands by the exponential enrichment (SELEX) technique. After 12 rounds of selection, 6 aptamers were selected and the corresponding binding affinities were assessed using fluorescence binding assays, revealing dissociation constants ranging from 15.45 to 40.98 nM. Notably, among the aptamers, RV2 and RV3 exhibited the highest binding affinities toward RVFV, with dissociation constants of 15.45 and 18.62 nM, respectively. Thiol-modified aptamers were subsequently immobilized onto screen-printed gold electrodes, facilitating the label-free detection of RVFV through square wave voltammetry. The voltammetric aptasensor demonstrated an excellent sensitivity, with a detection limit of 0.015 ng/mL. In addition, cross-reactivity assessments were conducted, where negligible response was obtained when the aptasensor was exposed to non-specific proteins.

The three members of the genus capripoxvirus (CaPV), lumpy skin disease virus (LSDV), sheeppox virus (SPPV), and goatpox virus (GTPV) have common hosts and areas of overlapping geographical distribution with Rift Valley fever virus (RVFV). Hence, to ensure more cost-effective disease surveillance we developed and evaluated a Luminex assay for the simultaneous detection of antibodies against CaPV and RVFV in domestic ruminants. In cattle, the assay had a sensitivity (Se) of 98.7% and a specificity (Sp) of 98.3% in detecting anti-LSDV antibodies; both diagnostic parameters were 100% for the detection of anti-RVFV antibodies in this species. In sheep and goats, Se and Sp were 100% for the detection of anti-SPPV and anti-GTPV antibodies while they were 100% and 98.9%, respectively for the detection of anti-RVFV antibody. The assay did not cross react with anti-parapoxvirus antibodies of cattle, sheep, and goats. This multiplex serological assay offers a practical tool for accurate detection and monitoring of the immunological status of domestic ruminant populations against veterinary and socio-economically important capripox- and phleboviral infections, thus has the potential to aid in the strategic application of vaccination programmes.

Rift Valley fever phlebovirus (RVFV, Phenuiviridae ) is an emerging arbovirus that can cause potentially fatal disease in many host species including ruminants and humans. Thus, tools to detect this pathogen within tissue samples from routine diagnostic investigations or for research purposes are of major interest. This study compares the immunohistological usefulness of several mono- and polyclonal antibodies against RVFV epitopes in tissue samples derived from natural hosts of epidemiologic importance (sheep), potentially virus transmitting insect species ( Culex quinquefasciatus, Aedes aegypti ) as well as scientific infection models (mouse, Drosophila melanogaster, C6/36 cell pellet). While the nucleoprotein was the epitope most prominently detected in mammal and mosquito tissue samples, fruit fly tissues showed expression of glycoproteins only. Antibodies against non-structural proteins exhibited single cell reactions in salivary glands of mosquitoes and the C6/36 cell pellet. However, as single antibodies exhibited a cross reactivity of varying degree in non-infected specimens, a careful interpretation of positive reactions and consideration of adequate controls remains of critical importance. The results suggest that primary antibodies directed against viral nucleoproteins and glycoproteins can facilitate RVFV detection in mammals and insects, respectively, and therefore will allow RVFV detection for diagnostic and research purposes.

Rift Valley Fever (RVF) is an emerging zoonotic arbovirus with a complex cycle of transmission that makes difficult the prediction of its expansion. Recent outbreaks outside Africa have led to rediscover the human disease but it remains poorly known. The wide spectrum of acute and delayed manifestations with potential unfavorable outcome much complicate the management of suspected cases and prediction of morbidity and mortality during an outbreak. We reviewed literature data on bio-clinical characteristics and treatments of RVF human illness. We identified gaps in the field and provided a practical algorithm to assist clinicians in the cases assessment, determination of setting of care and prolonged follow-up.

Diagnostic performance of an indirect enzyme-linked immunosorbent assay (I-ELISA) based on a recombinant nucleocapsid protein (rNP) of the Rift Valley fever virus (RVFV) was validated for the detection of the IgG antibody in sheep (n = 3367), goat (n = 2632), and cattle (n = 3819) sera. Validation data sets were dichotomized according to the results of a virus neutralization test in sera obtained from RVF-endemic (Burkina Faso, Democratic Republic of Congo, Mozambique, Senegal, Uganda, and Yemen) and RVF-free countries (France, Poland, and the USA). Cut-off values were defined using the two-graph receiver operating characteristic analysis. Estimates of the diagnostic specificity of the RVFV rNP I-ELISA in animals from RVF-endemic countries ranged from 98.6% (cattle) to 99.5% (sheep) while in those originating from RVF-free countries, they ranged from 97.7% (sheep) to 98.1% (goats). Estimates of the diagnostic sensitivity in ruminants from RVF-endemic countries ranged from 90.7% (cattle) to 100% (goats). The results of this large-scale international validation study demonstrate the high diagnostic accuracy of the RVFV rNP I-ELISA. Standard incubation and inactivation procedures evaluated did not have an adverse effect on the detectable levels of the anti-RVFV IgG in ruminant sera and thus, together with recombinant antigen-based I-ELISA, provide a simple, safe, and robust diagnostic platform that can be automated and carried out outside expensive bio-containment facilities. These advantages are particularly important for less-resourced countries where there is a need to accelerate and improve RVF surveillance and research on epidemiology as well as to advance disease control measures.

Following the Ebola outbreak in Western Africa in 2013–16, a global effort has taken place for preparedness for future outbreaks. As part of this response, the development of vaccines, treatments and diagnostic tools has been accelerated, especially towards pathogens listed as likely to cause an epidemic and for which there are no current treatments. Several of the priority pathogens identified by the World Health Organisation are haemorrhagic fever viruses. This review provides information on the role of reference materials as an enabling tool for the development and evaluation of assays, and ultimately vaccines and treatments. The types of standards available are described, along with how they can be applied for assay harmonisation through calibration as a relative potency to a common arbitrary unitage system (WHO International Unit). This assures that assay metrology is accurate and robust. We describe reference materials that have been or are being developed for haemorrhagic fever viruses and consider the issues surrounding their production, particularly that of biosafety where the viruses require specialised containment facilities. Finally, we advocate the use of reference materials at early stages, including research and development, as this helps produce reliable assays and can smooth the path to regulatory approval.

Rift Valley fever (RVF) is one of the main vector borne zoonotic diseases that affects a wide range of ruminants and human beings in Africa and the Arabian Peninsula. A rapid and specific test for RVF diagnosis at the site of a suspected outbreak is crucial for the implementation of control measures. A first-line lateral flow immunochromatographic strip test (LFT) was developed for the detection of the nucleoprotein (N) of the RVF virus (RVFV). Its diagnostic performance characteristics were evaluated using reference stocks isolates recovered from different hosts and in geographic regions mimicking clinical specimens and from known RVF negative serum samples. A high level of diagnostic accuracy (DSe (35/35), DSp (167/169)) was observed, including the absence of cross-reactivity with viruses belonging to different genera. The fact no specialized reagents and laboratory equipment are needed, make this assay a valuable, first-line diagnostic tool in resource-poor diagnostic territories for on-site RVFV detection, however the staff require training.

In southern African transfrontier conservation areas (TFCAs), people, livestock and wildlife share space and resources in semi-arid landscapes. One consequence of the coexistence of wild and domestic herbivores is the risk of pathogen transmission. This risk threatens local livelihoods relying on animal production, public health in the case of zoonoses, national economies in the context of transboundary animal diseases, and the success of integrated conservation and development initiatives. The level of interaction between sympatric wild and domestic hosts, defining different wildlife/livestock interfaces, characterizes opportunities of pathogen transmission between host populations. Exploring the relationship between infection burden and different types of wildlife/domestic interfaces is therefore necessary to manage the sanitary risk in animal populations through control options adapted to these multi-host systems. Here, we assessed the infection burdens of sympatric domestic cattle (Bos taurus/Bos indicus) and African buffalo (Syncerus caffer) at an unfenced interface and compared the infection burdens of cattle populations at different wildlife/livestock interfaces in the Great Limpopo TFCA. Patterns of infection in ungulate populations varied between wild and domestic hosts and between cattle populations at different wildlife/livestock interfaces. Foot-and-mouth disease, Rift Valley fever and theileriosis infections were detected in buffalo and cattle at unfenced interfaces; bovine tuberculosis was only present in buffalo; and brucellosis and lumpy skin disease only in cattle. At unfenced interfaces, cattle populations presented significantly higher Theileria parva and brucellosis prevalence. We hypothesize that cattle populations at wildlife/livestock interfaces face an increased risk of infection compared to those isolated from wildlife, and that the type of interface could influence the diversity and quantity of pathogens shared. Additional host behavioural and molecular epidemiological studies need to be conducted to support this hypothesis. If it is confirmed, the management of wildlife/livestock interfaces will need to be considered through the prism of livestock and public health.