IntroductionRabies is one of the major public health problems mostly affecting developing countries in Africa and Asia where 99.9% of all rabies related human deaths are recorded each year. In Democratic Republic of Congo, repeated outbreaks have been reported. Despite this, there is little reliable epidemiological data about rabies in the country for the development of effective control strategies.Materials and MethodsA retrospective study was carried out in Kinshasa Province during a period of five years (2009–2013) to describe the proportion of rabid animals and the species involved in rabies transmission and maintenance. The survey also aimed at describing the spatial-temporal distribution of rabies. To gather information, the daily registers of institutions involved in rabies diagnosis were reviewed and each rabies case was traced back to area of occurrence for collection of geographic coordinates.Results and DiscussionA total of 5,053 attacks were registered involving six animal species including dog, cat, monkey, rabbit, rat, and pig. Based on clinical observations, rabies was reported in dogs and cats while data obtained from the laboratory confirmed rabies cases included dogs, cats and a goat. The annual distribution showed a significant decrease of rabies cases from 2009 up to 2011 and a later increase up to 2013. There was no difference in rabies occurrence between seasons (p = 0.721). Rabies cases were three times higher in peri-urban zone than in urban zone OR = 3.4 (95% CI: 2.3–5.1). The positive proportion of rabies was 2.6% (95% CI: 2.1–3) based on clinical evidence and 65.9% (95% CI: 50–79.5) for laboratory confirmed cases.Conclusion and SuggestionThis study confirms the endemicity of rabies in Kinshasa where occurrence of rabies cases was related to human population density and lifestyle. In order to control rabies, there is need to set up a surveillance program and implement efficient mass vaccination campaigns of susceptible animals.

Crimean–Congo hemorrhagic fever virus (CCHFV) causes a zoonotic disease, Crimean–Congo hemorrhagic fever (CCHF) endemic in Africa, Asia, the Middle East, and Southeastern Europe. However, the prevalence of CCHF is not monitored in most of the endemic countries due to limited availability of diagnostic assays and biosafety regulations required for handling infectious CCHFV. In this study, we established a protocol to purify the recombinant CCHFV nucleoprotein (NP), which is antigenically highly conserved among multiple lineages/clades of CCHFVs and investigated its utility in an enzyme-linked immunosorbent assay (ELISA) to detect CCHFV-specific antibodies. The NP gene was cloned into the pCAGGS mammalian expression plasmid and human embryonic kidney 293 T cells were transfected with the plasmid. The expressed NP molecule was purified from the cell lysate using cesium-chloride gradient centrifugation. Purified NP was used as the antigen for the ELISA to detect anti-CCHFV IgG. Using the CCHFV NP-based ELISA, we efficiently detected CCHFV-specific IgG in anti-NP rabbit antiserum and CCHFV-infected monkey serum. When compared to the commercially available Blackbox CCHFV IgG ELISA kit, our assay showed equivalent performance in detecting CCHFV-specific IgG in human sera. These results demonstrate the usefulness of our CCHFV NP-based ELISA for seroepidemiological studies.

Crimean-Congo hemorrhagic fever virus (CCHFV), a nairovirus, is a tick-borne zoonotic virus that causes hemorrhagic fever in humans. The CCHFV nucleoprotein (NP) is the antigen most used for serological screening of CCHFV infection in animals and humans. To gain insights into antibody epitopes on the NP molecule, we produced recombinant chimeric NPs between CCHFV and Nairobi sheep disease virus (NSDV), which is another nairovirus, and tested rabbit and mouse antisera/immune ascites, anti-NP monoclonal antibodies, and CCHFV-infected animal/human sera for their reactivities to the NP antigens. We found that the amino acids at positions 161–320 might include dominant epitopes recognized by anti-CCHFV IgG antibodies, whereas cross-reactivity between anti-CCHFV and anti-NSDV antibodies was limited. Their binding capacities were further tested using a series of synthetic peptides whose sequences were derived from CCHFV NP. IgG antibodies in CCHFV-infected monkeys and patients were reactive to some of the synthetic peptide antigens (e.g., amino acid residues at positions 131–150 and 211–230). Only a few peptides were recognized by IgG antibodies in the anti-NSDV serum. These results provide useful information to improve NP-based antibody detection assays as well as antigen detection tests relying on anti-NP monoclonal antibodies.