anti-CCHFV IgG Research Articles

Exposure patterns and the risk factors of Crimean Congo hemorrhagic fever virus amongst humans, livestock and selected wild animals at the human/livestock/wildlife interface in Isiolo County, upper eastern Kenya.

Crimean Congo hemorrhagic fever (CCHF) is a tick-borne zoonotic disease caused by CCHF virus (CCHFV). The disease has a complex transmission cycle that involves a wide range of hosts including mammalian and some species of birds. We implemented a sero-epidemiological study in Isiolo County, Kenya, to determine relative seroprevalences of CCHFV in humans, livestock and in wild animals. In addition, we identified subject and environment level factors that could promote exposure to CCHFV. Humans (n = 580) and livestock (n = 2,137) were recruited into the study through a multistage random sampling technique, and in addition, various species of wild animals (n = 87) were also sampled conveniently. Serum samples from all recruited humans and animals were collected and screened for CCHFV antibodies using ID Screen multispecies, double-antigen IgG enzyme-linked immunosorbent assay (ELISA). The overall anti-CCHFV IgG seroprevalences in humans, cattle, goats, sheep and camels were 7.2% [95% CI: 3.1-15.8%], 53.9% [95% CI: 30.7-50.9%], 11.6% [95% CI: 7.2-22.5%], 8.6% [95% CI: 3-14%] and 89.7% [95% CI: 78-94%], respectively. On average, the sampled wild animals had CCHFV seroprevalence of 41.0% [95% CI: 29.1-49.4%]; giraffes had the highest mean CCHF seroprevalence followed by buffaloes, while impala had very low exposure levels. Statistical analyses using mixed effects logistic regression models showed that CCHFV exposure in humans was significantly associated with male gender, being over 30 years of age and belonging to a household with a seropositive herd. In livestock, a combination of animal- and environment level factors including older animals, being in an area with high normalized difference vegetation index (NDVI) and high vapour pressure deficit were significantly associated with CCHFV infection. Age, sex and species of wild animals were considered as the key risk factors in the analysis, but none of these variables was significant (P-value = 0.891, 0.401 and 0.664, respectively). Additionally, RT-qPCR analysis revealed the presence of CCHFV RNA in camels (30%), cattle (14.3%), and goats (3.8%), but not in humans, sheep, or wild animals. This study demonstrates that environmental factors, such as NDVI and vapor pressure deficit, affect CCHFV exposure in livestock, while the presence of infected livestock is the key determinant of human exposure at the household level. These findings underscore the importance of using One Health approaches to control the disease in human-livestock-wildlife interfaces. For instance, the existing CCHF surveillance measures could be enhanced by incorporating algorithms that simulate disease risk based on the environmental factors identified in the study. Additionally, tick control in livestock, such as the use of acaricides, could reduce CCHFV exposure in livestock and, consequently, in humans.

Seromolecular survey and risk factor analysis of Crimean-Congo haemorrhagic fever orthonairovirus in occupationally exposed herdsmen and unexposed febrile patients in Kwara State, Nigeria.

Crimean-Congo haemorrhagic fever virus (CCHFV) is a globally significant tick-borne zoonotic pathogen that causes fatal haemorrhagic disease in humans. Despite constituting an ongoing public health threat, limited research exists on the presence of CCHFV among herdsmen, an occupationally exposed population that has prolonged contact with ruminants and ticks. This cross-sectional study, conducted between October 2018 and February 2020 in Kwara State, Nigeria, was aimed at assessing CCHFV seroprevalence among herdsmen and non-herdsmen febrile patients, and identifying the associated risk factors. Blood samples from herdsmen (n = 91) and febrile patients in hospitals (n = 646) were analyzed for anti-CCHFV IgG antibodies and CCHFV S-segment RNA using ELISA and RT-PCR, respectively. Results revealed a remarkably high CCHFV seroprevalence of 92.3% (84/91) among herdsmen compared to 7.1% (46/646) in febrile patients. Occupational risk factors like animal and tick contact, tick bites, and hand crushing of ticks significantly contributed to higher seroprevalence in the herdsmen (p<0.0001). Herdsmen were 156.5 times more likely (p<0.0001) to be exposed to CCHFV than febrile patients. Notably, the odds of exposure were significantly higher (OR = 191.3; p<0.0001) in herdsmen with a history of tick bites. Although CCHFV genome was not detectable in the tested sera, our findings reveal that the virus is endemic among herdsmen in Kwara State, Nigeria. CCHFV should be considered as a probable cause of febrile illness among humans in the study area. Given the nomadic lifestyle of herdsmen, further investigations into CCHF epidemiology in this neglected population are crucial. This study enhances our understanding of CCHFV dynamics and emphasizes the need for targeted interventions in at-risk communities.

Evaluation of Nucleoprotein-Based Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Acute Crimean–Congo Hemorrhagic Fever Virus Infections in a Turkish Population

Background: Crimean-Congo hemorrhagic fever virus (CCHFV) causes a highly contagious tick-borne disease with high case-fatality rates in humans. It is circulating not only in many Asian and African countries, but also spreading to and within Europe. To cope better with future outbreaks of Crimean-Congo hemorrhagic fever (CCHF), the WHO has prioritized the need for the development and validation of CCHF diagnostics, including serological assays. In this study, we evaluated the performance of the new EUROIMMUN anti-CCHFV IgM and IgG enzyme-linked immunosorbent assays (ELISAs). Materials and Methods: Both ELISAs were compared to the Vector-Best VectoCrimean-CHF-IgM and -IgG ELISAs using the EUROIMMUN CCHFV Mosaic 2 IgM and IgG indirect immunofluorescence assays (IFA) as reference. Forty-nine acute-phase serum samples from patients with CCHFV infection confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and/or anti-CCHFV IgM IFA positivity were used to determine assay sensitivity. The assessment of specificity was based on sera from 30 control patients, 30 healthy blood donors, and 29 patients with hantavirus or sandfly fever virus infections. All samples originated from Turkey. Results: Sensitivity of the EUROIMMUN ELISAs (IgM 98.0%, IgG 47.1%) exceeded that of the Vector-Best ELISAs (IgM 95.9%, IgG 35.3%). Specificity of the EUROIMMUN ELISA IgM (86.4%) was slightly higher compared with the Vector-Best ELISA IgM (84.7%), while specificity for IgG was 100% for both assays. Qualitative agreement between the EUROIMMUN and Vector-Best ELISAs was substantial for detecting anti-CCHFV IgM (84.1%, ĸ = 0.673) and IgG (94.9%, ĸ = 0.791), whereas the quantitative results indicated a very strong positive correlation (IgM: r = 0.868, IgG: r = 0.913). Conclusion: The new EUROIMMUN anti-CCHFV ELISAs are standardized and easy-to-use tools that reliably support the identification of acute CCHF cases, and thus suitable for laboratories involved in on-site outbreak support.

Crimean-Congo Hemorrhagic Fever Virus Survey in Humans, Ticks, and Livestock in Agnam (Northeastern Senegal) from February 2021 to March 2022.

Crimean–Congo hemorrhagic fever virus (CCHFV) is widespread in Asia, Europe, and Africa. In Senegal, sporadic cases of CCHFV have been reported since 1960. Bordering Mauritania in northeastern Senegal, Agnam is an arid area in the region of Matam where CCHFV is endemic, which harbors a pastoralist community. Given the drought conditions of Agnam, inhabitants are in constant movement with their animals in search of pasture, which brings them into contact with pathogens such as arboviruses. To identify CCHFV in this area, we established a One Health site in order to analyze animal livestock, ticks and human samples collected over a one-year period by qRT-PCR and ELISA. Our analysis showed one (1/364) patient carried anti-CCHFV IgM and thirty-seven carried anti-CCHFV IgG (37/364). In livestock, anti-CCHFV IgG was detected in 13 (38.24%) of 34 sentinel sheep. The risk of CCHFV infection increased significatively with age in humans (p-value = 0.00117) and sheep (p-value = 1.18 × 10−11). Additional risk factors for CCHFV infection in sheep were dry seasons (p-value = 0.004) and time of exposure (p-value = 0.007). Furthermore, we detected a total of three samples with CCHFV RNA within Rhipicephalus evertsi evertsi and Rhipicephalus guilhoni tick species. Our results highlighted the usefulness of a One Health survey of CCHFV in pastoral communities at risk of arboviruses.

Crimean-Congo hemorrhagic fever virus circulating among sheep of Portugal: a nationwide serosurvey assessment.

Crimean-Congo hemorrhagic fever virus (CCHFV) is a widespread zoonotic pathogen that can cause mild to severe hemorrhagic disease in humans. CCHFV may be transmitted through direct contact with tissue or blood of viremic animals; however, the primary transmission route is through infected tick bites. CCHFV RNA has been detected in ticks feeding on domestic and wild animals in western Spain, suggesting an established circulation of CCHFV in Western Europe. Ruminants have been recognized as important CCHFV reservoirs and have been linked to human cases in endemic regions. Given the emergence of CCHF in neighboring Spain, and a report of two CCHFV seropositive humans in southern Portugal in 1985, we investigated the potential circulation of this virus in the country by performing a nationwide anti-CCHFV IgG serosurvey in sentinel sheep of Portugal. Sera (n = 459) randomly selected from widely distributed farms (n = 20) of Portugal were tested using a commercial double-antigen enzyme-linked immunosorbent assay, yielding an overall seroprevalence of 0.4% (95% confidence interval [CI] 0.04-1.56%). Positive sheep were from the southern region of Portugal (Alentejo region), which raise the seroprevalence of this region to 0.74% (95% CI 0.09-2.66%). This is the first study reporting the presence of CCHFV antibodies in sheep of Portugal, thus suggesting a geographical expansion of CCHFV to this country. It seems likely that CCHFV may exist focally in southern Portugal.

Mapping of Antibody Epitopes on the Crimean-Congo Hemorrhagic Fever Virus Nucleoprotein

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.

Evcil ruminatlarda Kırım-Kongo Kanamalı Ateşi Virusu (KKKAV) enfeksiyonunun seroepidemiyolojik araştırması, Adana ili, Doğu Akdeniz bölgesi, Türkiye.

Crimean-Congo hemorrhagic fever virus (CCHFV) has been observing since 2002 and 2015 in Turkey and Adana, respectively. However, there were no previous seroepidemiological data on CCHFV in Adana. This study, thus, aimed to investigate the seroepidemiological status of Adana province for CCHFV in ruminants, according to species, breed, age, gender and geographical location. The study was carried out in 15 districts from sea level to plateaus (0-1472 m), within a 14,030 km² area. 485 samples were tested with modified anti-CCHFV IgG ELISA. A human-based IgG ELISA was adapted to recognize cattle, sheep and goats IgG antibodies. The conjugation (anti-human IgG peroxidase) was replaced with appropriate conjugate in each species. Statistical analyses indicated a significant relationship between antibody presence and animal age, species, breed and location but there was no relation with animal gender. Out of 485 samples, 154 were seropositive (31.8%). It was determined that the statistical cluster varied between 5.8% and 74.3% according to location and seropositivity. The most important variable associated with antibody presence was geographic location. While high seropositivity is found at altitudes above 555 meters, it is found to be 7% and 3% at sea level. The seropositivity reached 100% in the village where human deaths were seen in 2015, and 74.3% in the Tufanbeyli district where this village is located. The results indicate that Adana is an endemic province for CCHFV infection, especially in high altitude regions.

Sequential determination of viral load, humoral responses and phylogenetic analysis in fatal and non-fatal cases of Crimean-Congo hemorrhagic fever patients from Gujarat, India, 2019.

BackgroundThirty-four CCHF cases (17 fatal; 17 survived) were confirmed from Gujarat state, India during the year 2019. We aimed to find out the viral load, antibody kinetics, cytokine profile and phylogenetic analysis between fatal and non- fatal cases.MethodsThirty four cases were included in this study. Blood and urine samples were collected from all the cases on the day of admission to hospital. Non-fatal cases were followed weekly for understanding the profile of viral kinetics, anti-CCHFV IgM and IgG antibodies. We also quantified the cytokines in both fatal and non-fatal cases. For epidemiological correlation, livestock were screened for anti-CCHF IgG antibodies and the tick pool specimens were tested by real time RT-PCR. Virus isolation was attempted on tick pools and human specimens and phylogenetic analysis performed on human and ticks complete genome sequences.ResultsCCHF cases were detected throughout year in 2019 with the peak in August. Out of 34 cases, eight secondary CCHF cases were reported. Cases were predominantly detected in males and in 19–45 years age group (55.88%). The persistence of viremia was observed till 76th POD (post onset date) in one case whereas anti-CCHFV IgM and IgG was detected amongst these cases from the 2nd and 20th POD respectively. Positivity observed amongst livestock and tick pools were was 21.57% and 7.4% respectively. The cytokine analysis revealed a significant increase in the level of serum IL-6, IL-10 and IFN-γ during the acute phase of the infection, but interestingly IL-10 lowered to normal upon clearance of the virus in the clinically recovered case. Fatal cases had high viral RNA copy numbers. Bleeding from one or two mucosal sites was significantly associated with fatality (OR-16.47;p-0.0034 at 95% CI). We could do CCHF virus isolation from two cases. Phylogenetic analysis revealed circulation of re-assortment of Asian-West African genotypes in humans and ticks.ConclusionsThe persistence of CCHF viral RNA was detected till 76th POD in one of the survivors. The circulation of a re-assortment Asian-West African genotype in a CCHF case is also reported first time from India.

Determination of Seroprevalence and Risk Factors of Crimean-Congo Haemorrhagic Fever (CCHF) in the Endemic Region in Turkey: A Population-Based Cross-Sectional Study.

Objectives Turkey is one of the countries that has the most cases of CCHF in recent years among the endemic countries. The disease also poses an important health threat with high mortality rate. The aim of the study was to determine the seroprevalence and risk factors of CCHF in adults aged ≥20 years in Tokat in the endemic region, Turkey. Methods In this population-based cross-sectional study, a total of 85 Family Medicine Units (FMUs), from over 170 in Tokat, were randomly selected using 50% sampling. The sample size was determined among the subjects aged ≥20 who registered with the FMUs, due to gender, age group, and the urban/rural population size of Tokat using the stratified cluster sampling method. Subjects were invited to the FMUs. A questionnaire was performed face to face. The blood samples were taken, and anti-CCHFV IgG antibodies were measured with ELISA method. Results 1272 (54.9%) out of 2319 participants were female, and the mean age was 47.3 ± 15.3. Anti-CCHFV IgG seropositivity was 5.6% (n = 130). Seropositivity rates in terms of adjusted odds ratios (AOR) were higher 2.53 times (95% CI: 1.57–4.08; p=0.001) in males; 4.05 (95% CI: 2.14–7.65; p < 0.001) in age group ≥65; 0.33 (95% CI: 0.14–0.76; p < 0.001) in graduates of high school and above; 0.71 (95%CI: 0.33–1.52; p < 0.001) in ones with good income; 1.84 (95%CI: 1.18–2.86; p < 0.001) in farmers; 1.64 (95% CI: 1.04–2.27; p < 0.001) in people dealing with animal husbandry; and 1.02 (95% CI: 1.03–2.29; p < 0.001) in those with history of tick contact. Conclusions CCHF seroprevalence is still a common public health problem in Tokat, Turkey. Male gender, advanced age group, low-educated, low-income, farmers, animal husbandry, and history of tick contact were found to be risk factors for CCHF. The importance of this kind of community-based studies to identify the seroprevalence in regional and national level increases even more.

New circulation of genotype V of Crimean-Congo haemorrhagic fever virus in humans from Spain.

Crimean-Congo haemorrhagic fever (CCHF) is a widespread tick-borne viral disease caused by the Crimean-Congo haemorrhagic fever virus (CCHFV). CCHFV has been implicated in severe viral haemorrhagic fever outbreaks. During the summer of 2016, the first two cases with genotype III (Africa 3) were reported in Spain. The first objective of our study was to determine the presence of CCHFV among patients with febrile illness during the spring and summer periods in 2017 and 2018. Finally, we perform a phylogenetic analysis to determine the genotype of the virus. We prospectively evaluated patients aged 18 years and older who came to the emergency department at the Salamanca's University Hospital (HUS) with fever. Specific IgM and IgG antibodies against CCHFV by ELISA and one immunofluorescence assay against two different proteins (nucleoprotein and glycoprotein C) was done. Moreover, molecular detection by Real Time PCR was performed in all collected samples. A phylogenetic analysis was carried out to genetically characterize CCHFV detected in this study. A total of 133 patients were selected. The mean age was 67.63 years and 60.9% were male. One-third of the patients presented an acute undifferentiated febrile illness. Three patients had anti-CCHFV IgG antibodies, suggesting a previous infection. One patient had anti-CCHFV IgM antibodies and a confirmatory RT-PCR. Phylogenetic analysis indicated that the virus corresponds to the European genotype V. This patient came to the emergency department at HUS in August 2018 presenting an acute febrile syndrome with thrombopenia and liver impairment. We describe a new circulation of European genotype V CCHFV in Spain. Moreover, this study suggests that CCHFV is an identifiable cause of febrile illness of unknown origin in Spain. Thus, CCHF could be suspected in patients with fever, liver damage, and/or haemorrhagic disorders, particularly in people with risk activities who present in the spring or summer.

Purification of Crimean\u2013Congo hemorrhagic fever virus nucleoprotein and its utility for serological diagnosis

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.

Absence of IgG antibodies among high-risk contacts of two confirmed cases of Crimean-Congo haemorrhagic fever in the autonomous region of Madrid (Spain)

Crimean-Congo haemorrhagic fever (CCHF) is a widely distributed tick-borne disease. In Spain, the disease has emerged as outbreak associated with high-risk exposures. Our goal was to evaluate the prevalence of antibodies against the CCHF virus (CCHFV) in high-risk contacts. A cross-sectional study was conducted. Three hundred eighty-six high-risk contacts were identified comprising family contacts and hospital workers who had attended the cases. Fifty-seven cases with closer exposure were selected. However, forty-nine cases participated in the study. IgG antibodies were detected by immunoenzymatic techniques. All determinations tested negative for anti-CCHFV IgG antibodies. Most of the responders were women (73.5%), and belong to the intensive care department (53.1%). In relation to other possible sources of exposures, 18.4% travelled to countries with CCHF transmission risk. No CCHF positivity was recorded among selected high-risk contacts. This highlights the importance of standard precautions which might have protected healthcare workers and care providers from CCHF infection.

Seroprevalence of Crimean‐Congo haemorrhagic fever among three selected risk human groups in disease‐endemic region of Pakistan

The occurrence of Crimean-Congo haemorrhagic fever (CCHF) in humans is linked with animals living in close vicinity, and information on the incidence of CCHF at the human-animal interface is scarce. Therefore, the current study was designed to identify the high-risk groups of individuals linked with animals in the Chakwal district of Pakistan having a history of CCHF cases in humans. In subject matter, coupled with risk factor analysis, we performed a sero-based CCHF surveillance in three selected risk groups of humans including abattoir workers (n=137), milkmen (n=169) and animal handlers (n=147). Sera samples and questionnaire-based data were collected from each of the participants and screened for anti-CCHFV IgG antibodies using enzyme-linked immunosorbent assay. The highest seroprevalence was observed in animal handlers (n=14, 9.52%, 95% CI: 4.68-13.99) followed by abattoir workers (n=9, 6.57%, 95% CI: 2.42-10.72) and milkmen (n=3, 1.78%, 95% CI: 0.24-4.24). The risk of seropositivity was significantly associated with humans linked with tick-infested animals (OR: 11.0, 95% CI: 1.5-83.0, p=.002), old age >40years (OR: 6.6, 95% CI: 2.7-16.0, p<.0001), illiteracy (OR: 4.3, 95% CI: 1.5-13.0, p=.004) and humans without knowledge about CCHF (OR: 7.6, 95% CI: 1.8-33.0, p=.0009). The findings of the current study highlighted the seroprevalence of CCHF in high-risk groups of humans living in a disease-endemic area of Pakistan and highlight the need for well-integrated disease surveillance in the future to better comprehend disease control interventions.

Crimean-Congo haemorrhagic fever (CCHF) virus-specific antibody detection in blood donors, Castile-León, Spain, summer 2017 and 2018.

BackgroundCrimean-Congo haemorrhagic fever virus (CCHFV) is considered an emerging or even a probable re-emerging pathogen in southern Europe. Presence of this virus had been reported previously in Spain in 2010.AimWe aimed to evaluate the potential circulation of CCHFV in western Spain with a serosurvey in asymptomatic adults (blood donors).MethodsDuring 2017 and 2018, we conducted a CCHFV serosurvey in randomly selected asymptomatic blood donors from western Spain. Three assays using specific IgG antibodies against CCHFV were performed: the VectoCrimea ELISA test, an in-house ELISA and indirect immunofluorescence (EuroImmun) test with glycoprotein and nucleoprotein.ResultsA total of 516 blood donors participated in this cross-sectional study. The majority of the study participants were male (68.4%), and the mean age was 46.3 years. Most of the participants came from rural areas (86.8%) and 68.6% had contact with animals and 20.9% had animal husbandry practices. One in five participants (109/516, 21.1%) were engaged in at-risk professional activities such as agriculture and shepherding, slaughtering, hunting, veterinary and healthcare work (mainly nursing staff and laboratory technicians). A total of 15.3% of the participants were bitten by ticks in the days or months before the date of sampling. We detected anti-CCHFV IgG antibodies with two diagnostic assays in three of the 516 individuals and with one diagnostic assay in six of the 516 individuals.ConclusionSeroprevalence of CCHFV was between 0.58% and 1.16% in Castile-León, Spain. This is the first study in western Spain that showed circulation of CCHFV in healthy people.

Crimean Congo hemorrhagic fever serosurvey in humans for identifying high-risk populations and high-risk areas in the endemic state of Gujarat, India

BackgroundCrimean Congo Hemorrhagic Fever (CCHF) is a highly infectious zoonotic disease of humans transmitted by Hyalomma ticks. Earlier studies have shown CCHF seroprevalence in livestock throughout India, yet sporadic outbreaks have been recorded mostly from the Gujarat state of India since 2011. Occupational vulnerability to CCHF for animal handlers, veterinarians, abattoir workers, and healthcare workers has been documented. The current study was planned to determine the seroprevalence of CCHF with an intention to identify the high -risk population and high -risk areas from Gujarat state, India.MethodsBased on the socio-clinical data, the human population of Gujarat was divided into eight categories viz. A: CCHF affected person/house/close contact, B: Neighborhood contacts, C: Animal handlers, D: General population, E: Farmers, F: Abattoir workers, G: Veterinarian, H: Healthcare workers. A total of 4978 human serum samples were collected from 33 districts of Gujarat during year 2015, 2016 and 2017. All the samples were screened for the presence of anti-CCHFV IgG using indigenously developed anti-CCHFV IgG ELISA. Univariate regression analysis was performed to recognize significant risk factors for CCHF seropositivity.ResultsTwenty-five serum samples were found to be positive with an overall CCHF human seropositivity of 0.5% (95% CI 0.30–0.74%). Gender predisposition to CCHF prevalence was observed in males (OR: 2.80; p-value: 0.020). The risk for seropositivity increased sevenfold when a person was in contact or neighbor with a CCHF case (OR 7.02; p-value: < 0.0001). No significant difference in seropositivity was observed within different age groups. Veterinarians, healthcare workers, and control group were found to be seronegative for CCHF.ConclusionsIn-spite of CCHF sporadic outbreaks reported in Gujarat, the seropositivity for CCHF in the state was low as compared to other endemic countries. Males, close contacts and neighbors were identified as a high-risk population for CCHF infection. To recognize the high-risk area, tick screening and animal serosurvey would be a wiser choice. The study also suggests circulation and under diagnoses of CCHFV in the naïve regions of Gujarat.

Seroepidemiology of CCHF in Domestic Animals in Endemic Areas in Albania

Abstract Crimean-Congo hemorrhagic fever (CCHF) is an arboviral zoonotic infection wich is endemic in some areas of the country. The aim of this study was to assess the seroprevalence of CCHFV in previous and recent endemic areas of the country. This crosssectional serologic study was conducted in period 2010-2014 by the Institute of Public health in Tirana, Albania. The survey included 152 sera specimen of cattle which were randomly collected from seven districts in Albania over the period 2010-2014. All samples were collected randomly from seven districts in Albania and were examined for anti-CCHFV IgG. Of the total of 152 samples examined, 102 (67.1%) were positive to IgG ELISA. The highest positivity rate was among cows (88.3%) and in recent endemic areas of Has and Kukes, 89.7% and 82.9% respectively. In recent endemic areas humans can potentially contract the disease as compared to other areas of the country.

First serological evidence of Crimean-Congo haemorrhagic fever in febrile patients in Mozambique

ObjectiveDespite its geographical spread, the epidemiology of Crimean-Congo haemorrhagic fever (CCHF) in Sub-Saharan Africa is incompletely understood and its occurrence in Mozambique is unknown. This study was conducted with the aim of investigating the occurrence of CCHF virus (CCHFV) among febrile patients attending an outpatient appointment clinic at three separate primary health care centres in Mozambique. MethodsSerum samples were collected from a total of 300 febrile patients aged >5 years who were recruited between March 2015 and March 2016 at three health centres in Mozambique. Each patient was screened for IgG antibodies against CCHFV using an ELISA. ResultsOf the 300 patients enrolled, eight had samples that were positive for anti-CCHFV IgG antibodies, yielding a prevalence rate of 2.7%. ConclusionsThis study shows for the first time that humans are exposed to CCHFV in Mozambique. It highlights the need for further work to investigate the broader extent of circulating CCHFV in the country and its clinical implications.

Plant-produced Crimean-Congo haemorrhagic fever virus nucleoprotein for use in indirect ELISA

Crimean-Congo haemorrhagic fever (CCHF) is a disease of serious public concern caused by the CCHF virus (CCHFV). Anti-CCHFV IgG in humans can be detected using ELISA with native antigen prepared from cell cultures which have been infected with virus or from brain tissue of suckling mice which have been inoculated with virus. However, the preparation of these reagents requires high biosafety levels and is expensive. A safer, more cost-effective recombinantly-produced NP reagent is desirable. Recently, plants have been shown to be a cost-effective and safe system for expression of recombinant proteins. This work describes cloning of the CCHFV NP gene into three different plant expression systems and comparison of expression in Nicotiana benthamiana. The highest expressing construct was selected. Expressed NP was purified by ammonium sulphate fractionation prior to histidine affinity chromatography. Purified NP was tested in an indirect ELISA to determine if the recombinant antigen was able to detect anti-CCHFV IgG in sera from convalescent patients. Plant-produced NP detected IgG antibodies against CCHFV in 13/13 serum samples from convalescent patients and 0/13 samples collected from volunteers with no history of CCHFV infection. Results were compared with commercially available immunofluorescent assays and 100% concordance was obtained between the two assays. This suggests that a full evaluation of the plant produced NP for application as a safe recombinant is warranted.

Seroprevalence of Crimean-Congo Hemorrhagic Fever in Turkey's Van Province.

Crimean-Congo hemorrhagic fever (CCHF) is an endemic tick-borne viral disease that affects both animals and humans. This study aims to determine the seroprevalence of CCHF in Turkey's Van province using analysis of blood samples obtained from people living in the region. Blood specimens were taken from healthy subjects living in Van province and some of the surrounding villages between January and July 2012. Blood samples were initially tested using a CCHF virus (CCHFV) IgM IgG kit for anti-CCHFV IgG, followed by anti-CCHFV IgM determination of any IgG positive blood samples. IgM-positive specimens were re-confirmed using real-time polymerase chain reaction (qPCR). One hundred and 7 men and 261 women were included in the study. Fifty-three blood specimens (14.4%) were anti-CCHFV IgG positive, and 2 of these were anti-CCHFV IgM positive. Two blood samples with anti-CCHFV IgM seropositivity tested negative using qPCR, indicating chronic infections. Locality, sex, and a history of tick bites did not significantly affect anti-CCHFV IgG seropositivity. Although the incidence of anti-CCHFV IgG in blood specimens was 14.4%, no deaths have yet been reported in Turkey's Van province. It is imperative that clinical CCHFV tests be implemented for people at high risk of developing CCHFV-related complications.

Crimean-Congo Hemorrhagic Fever Virus IgG in Goats, Bhutan

To The Editor: Crimean-Congo hemorrhagic fever (CCHF) is a highly infectious tickborne disease caused by a high-risk group of viruses belonging to the family Bunyaviridae (1,2). In humans, the overall case-fatality rate of CCHF is ≈30%, but in severe and hospitalized patients, fatalities may be up to 80% (3,4). CCHF is widespread in various countries in Africa, Asia, and Europe; the virus had been identified in humans in China, Pakistan, and Afghanistan and has been recently reported for the first time in humans in India (4–7). Humans can be infected by bites from infected ticks, mainly of the Hyalomma genus; by unprotected contact with blood or tissue of viremic patients; or during slaughtering of infected animals. In addition, nosocomial infections are found in humans (1,4,8,9). Fatal cases of CCHF in humans were confirmed in Ahmadabad in India in 2011, but a recent serosurvey in livestock showed that this disease has widespread seroprevalence in domestic animals across India (7–10). Bhutan shares a long, porous border with India, and animals and humans frequently cross the border. Comprehensive surveillance was needed to determine the presence of CCHF virus (CCHFV) in livestock in Bhutan and to assess risk for zoonotic infection in humans. During October 2015, in collaboration with the National Centre for Animal Health Bhutan, we retrospectively tested serum samples collected during April–May 2015 from 81 goats and 92 cattle for CCHFV-specific IgG by using ELISA kits (Sheep/goat anti-CCHFV IgG ELISA kit and Cattle anti-CCHFV IgG ELISA kit; National Institute of Virology, Pune, India), as described (10). CCHFV IgG was detected in 31 (38.2%) goats; no cattle had positive results. The samples from goats, which were collected in early 2015 as part of surveillance of peste des petits ruminants, originated from the southern district of Sarpang, which shares a porous border with the state of Assam in India (Figure). The samples from cattle were collected from the National Nublang Breeding Center (Trashigang district) and the National Jersey Breeding Center (Samtse district) (Figure). Findings indicated that all goats that tested positive for CCHFV were reported to have been either bred within households that kept goat herds or procured from other villages within the district. Exact sources of those seropositive goats could not be ascertained. However, in a few instances in the past, breeding goats (male and female) were procured from India by the Bhutan government and distributed to farmers for breed improvement. We also believe that cross-border movement of animals and unofficial imports of goats by farmers along the porous borders of southern Bhutan likely occurred. Furthermore, a large number of dairy cattle are imported annually from India for enhancing milk production and breeding purposes. Not all imported animals (both cattle and goats) were tested for CCHF because of a lack of diagnostic facilities and the negligible occurrence of the disease in livestock. Figure Locations in Bhutan where serum samples were collected from goats (triangles) and cattle (square and circle) and tested for Crimean-Congo hemorrhagic fever virus. The shaded area shows the boundaries of Sarpang district and subdistricts, where samples ... Our findings indicate that the risk of importing emerging infectious diseases along with live animals poses a serious risk to public health. Consequently, detailed risk-based surveillance is necessary to understand the complete scenario of CCHFV prevalence in livestock in Bhutan because Hyalomma tick species, the primary vectors of CCHF, are present on animals here. In addition, a survey among at-risk human populations is also needed. Findings from these surveillance activities would help institute more diagnostic facilities and risk-based surveillance and assist in developing a preparedness plan at the human–animal interface. Although our study has limitations because of the low number of serum samples tested from limited animal species from only 3 areas, the study provides evidence that CCHFV is circulating in goats in Bhutan.