Immature platelet fraction as a useful marker in Crimean-Congo hemorrhagic fever

@#The aim of this study is to evaluate the clinical significance and diagnostic performance of the immature platelet fraction (%IPF) in Crimean-Congo hemorrhagic fever (CCHF). Samples obtained from 32 healthy control subjects and 40 CCHF patients (9 positive and 31 negative radiological findings) were evaluated in the study. The samples obtained from CT-positive subjects demonstrated higher IPF% values which also exhibited a positive correlation with mean platelet volume (MPV) and platelet size deviation width (PDW) values.The patient group IPF% values were positively correlated with the duration of hospital stay. The ROC analysis also suggested the potential importance of IPF values higher than 10.5% in diagnosing CCHF patients with positive radiological findings.The results of our study showed that % IPF can be considered as a useful parameter in the follow-up of the disease course in patients with CCHF.

Molecular detection of Crimean-Congo Haemorrhagic Fever (CCHF) virus in hard ticks from South Khorasan, east of Iran

Background & objectives: Crimean-Congo Hemorrhagic Fever (CCHF) is a deadly viral infection reported from more than 30 countries. It is considered a zoonosis? and tick bites are the main route of transmission in nature. So far, the virus has been identified in 31 species of hard (Ixodidae) and soft (Argasidae) ticks. The aim of this study was to determine the rate of CCHF virus infection in hard ticks from South-Khorasan province, east of Iran. Methods: In this study, 684 livestock including 302 sheep, 344 goats, 16 cows and 22 camels were sampled from Birjand, Qaen, Khusf, Darmian and Sarbisheh counties. Genus and species of the ticks were diagnosed under stereomicroscope according to valid morphological keys. Reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect the CCHF virus genome based on S segment in 100 ticks. Results: RT-PCR detected CCHF virus genome in 7 out of 100 ticks. Positive ticks belonged to Hyalomma and Rhipicephalus genera. CCHF virus infected species were Rhipicephalus sanguineus, Hyalomma detritium and Hyalomma asiaticum. All the infected ticks were isolated from goat and sheep and were from Birjand county. Interpretation & conclusion: Our results suggest that Hyalomma and Rhipicephalus may be the main vectors of CCHF virus in the study area.

Role of TGF-?1 and PDGF-B in Crimean-Congo hemorrhagic fever in Eastern Anatolia Region in Turkey

Background & objectives: In this study, we aimed to investigate the relationship between serum TGF-?1 and PDGFB levels with the pathogenesis, clinical course and prognosis of adult Crimean-Congo hemorrhagic fever (CCHF) patients. Methods: 50 adult patients and 30 healthy individuals as a control group were included in the study, who were followed up and treated with the diagnosis of CCHF at the Atatürk University Faculty of Medicine Infectious Diseases and Clinical Microbiology Clinic, between March 2017 and September 2019 in Eastern Anatolia Region in Turkey. Blood samples were taken from patients on the first day of their hospitalization and on the sixth day of their complaints. TGF-?1 and serum PDGF-B levels were studied by ELISA method using commercial kits, from serum samples taken from CCHF patient group and individuals in healthy control group and stored at -80°C. Results: While the serum TGF- ?1 levels of patients with CCHF were found to be significantly higher on the sixth day of their complaints compared to the first day of hospitalization (42.33 ± 15.42, 28.40 ± 7.06, p = 0.001, respectively), the serum PGDF-B levels were found to be significantly lower on the sixth day of their complaints compared to those measured on the day of hospitalization (62.14 ± 19.75, 93.96 ± 20.02, respectively, p = 0.001). Interpretation & conclusion: Serum TGF-?1 levels are higher and PDGF-B levels are lower in CCHF patients with severe disease, indicating that serum TGF-?1 and PDGF-B play an important role in the pathogenesis of CCHF

Significance of IL-1Ra and IL-6 gene variants in Turkish patients with Crimean-Congo hemorrhagic fever

Objective: To investigate the association between IL-1Ra variable number of tandem repeat (rs2234663), IL-6 -597GA (rs1800797), IL-6 -572GC (rs1800796) and the risk of Crimean-Congo hemorrhagic fever (CCHF) in the Turkish patients. Methods: This study included 50 patients infected with CCHF and 50 healthy controls. These variants were genotyped using polymerase chain reaction and/or restriction fragment length polymorphism method. Results: The distribution of the IL-6 -572GC genotypes and alleles varied significantly between the patients and the controls. The subjects carrying IL-6 -572GC GG genotype and G allele had increased risk of developing CCHF compared to the control group (P=0.006, P=0.014, respectively). IL-6 -572GC GC genotype was higher in the controls than the patients (P=0.006). For the triple genotype combinations, the 1/2-GC-GG genotype combination was detected more frequently in the control group than CCHF patients (P=0.016). IL-6 (-572/-597) GG-GG genotype was significantly higher in the patient group (P=0.015), while the GC-GG genotype was significantly lower in the patient group (P=0.005). Additionally, the G-G haplotype was significantly higher in the patient group (P=0.042), whereas C-G was found to be significantly lower in the patients than the control group (P=0.037). Conclusions: The results of this study suggest the IL-6 -572GC variant might be genetic markers of sensitivity to CCHF in the Turkish population and may facilitate greater protection against the disease.

Significance of IL-1Ra and IL-6 gene variants in Turkish patients with Crimean-Congo hemorrhagic fever

Objective: To investigate the association between IL-1Ra variable number of tandem repeat (rs2234663), IL-6 -597GA (rs1800797), IL-6 -572GC (rs1800796) and the risk of Crimean-Congo hemorrhagic fever (CCHF) in the Turkish patients. Methods: This study included 50 patients infected with CCHF and 50 healthy controls. These variants were genotyped using polymerase chain reaction and/or restriction fragment length polymorphism method. Results:The distribution of the IL-6 -572GC genotypes and alleles varied significantly between the patients and the controls. The subjects carrying IL-6 -572GC GG genotype and G allele had increased risk of developing CCHF compared to the control group (P=0.006, P=0.014, respectively). IL-6 -572GC GC genotype was higher in the controls than the patients (P=0.006). For the triple genotype combinations, the 1/2-GC-GG genotype combination was detected more frequently in the control group than CCHF patients (P=0.016). IL-6 (-572/-597) GG-GG genotype was significantly higher in the patient group (P=0.015), while the GC-GG genotype was significantly lower in the patient group (P=0.005). Additionally, the G-G haplotype was significantly higher in the patient group (P=0.042), whereas C-G was found to be significantly lower in the patients than the control group (P=0.037). Conclusions: The results of this study suggest the IL-6 -572GC variant might be genetic markers of sensitivity to CCHF in the Turkish population and may facilitate greater protection against the disease.

Recombinant Crimean-Congo hemorrhagic fever virus with Nanoluciferase reporter and its potential application for rapid antiviral compounds screening / 中华实验和临床病毒学杂志

Objective@#To construct the recombinant Crimean-Congo hemorrhagic fever virus (CCHFV) which can express the secreted nanoluciferase (NanoLuc) and investigate its potential application for rapid antiviral compounds screening.@*Methods@#The ORF of NanoLuc and the mucin encoded by the M segment of CCHFV were merged, and the recombinant CCHFV (rCCHFV) was rescued through reverse genetic system. Then rCCHFV was used to evaluate the antiviral effect for ribavirin and Furin inhibitor in vitro.@*Results@#The rCCHFV_mucin_NLuc with NanoLuc reporter was obtained, and the relative light unit (RLU) which can reflect NanoLuc activity was positively correlated with median tissue culture infective dose (TCID50) in the infected cell supernatant (cor=0.998, P=0.001). When the concentration for the compounds was 10 μmol/L, there was no significant difference for the NanoLuc activity in the infected cell supernatant between Furin inhibitor and ribavirin (P > 0.1) from day 1 to 3 after treatment. But at day 4, the NanoLuc activity in Fruin inhibitor treated group was significantly higher than that of ribavirin treated group (P=0.001), and no significant difference was found between the Furin inhibitor and untreated group (P > 0.1).@*Conclusions@#The rCCHFV with NanoLuc reporter was recovered successfully and it could be used for the primary rapid screening of antiviral compounds in future.

Knowledge, attitude and practice of healthcare workers concerning Crimean-Congo hemorrhagic fever in Western Iran

Objective: To determine the knowledge, attitude and practice of healthcare workers in Kermanshah Province about Crimean-Congo hemorrhagic fever (CCHF). Methods: This study was conducted in 2014 on healthcare personnel in different job categories including physicians, nurses, midwives, laboratory staff and network health staff of Kermanshah Province by direct interview. Results: A total of 367 respondents who had more than 5 years of experience in their jobs were interviewed. Among them 91% of physicians and nurses, 97% of midwives and health workers and 96% of laboratory staff stated that they had not been confronted with CCHF patients so far. Regarding knowledge, 76% of physicians, 78% of nurses, 77% of midwives and 58% of laboratory staff believed that the disease is remediable. Most of the interviewed participants stated that the disease pertains to people who are in close contact with domestic animals, but they did not consider their own occupations as one of the risk factors. More than 70% of the respondents believed that the disease may exist in the province or their work field. Generally, the knowledge about CCHF was inadequate, with nurses having the lowest level of knowledge. Conclusions: Knowledge of Kermanshah healthcare staff about CCHF was poor, especially nurses in a high risk job category. Therefore, it is necessary to conduct specific training programs for the disease identification, transmission, prevention, and treatment as well as the use of personal protection and safety devices.

Knowledge, attitude and practice of healthcare workers concerning Crimean-Congo hemorrhagic fever in Western Iran

Objective: To determine the knowledge, attitude and practice of healthcare workers in Kermanshah Province about Crimean-Congo hemorrhagic fever (CCHF). Methods: This study was conducted in 2014 on healthcare personnel in different job categories including physicians, nurses, midwives, laboratory staff and network health staff of Kermanshah Province by direct interview. Results: A total of 367 respondents who had more than 5 years of experience in their jobs were interviewed. Among them 91%of physicians and nurses, 97%of midwives and health workers and 96%of laboratory staff stated that they had not been confronted with CCHF patients so far. Regarding knowledge, 76%of physicians, 78%of nurses, 77%of midwives and 58%of laboratory staff believed that the disease is remediable. Most of the interviewed participants stated that the disease pertains to people who are in close contact with domestic animals, but they did not consider their own occupations as one of the risk factors. More than 70% of the respondents believed that the disease may exist in the province or their work field. Generally, the knowledge about CCHF was inadequate, with nurses having the lowest level of knowledge. Conclusions: Knowledge of Kermanshah healthcare staff about CCHF was poor, especially nurses in a high risk job category. Therefore, it is necessary to conduct specific training programs for the disease identification, transmission, prevention, and treatment as well as the use of personal protection and safety devices.

Evaluation of the Crimean-Congo Hemorrhagic Fever Cases Followed and Treated in Our Clinic between 2009-2013.

Objectives: The Crimean-Congo hemorrhagic fever virus (CCHFV) belongs to the family Bunyaviridae, genus Nairovirus, and causes severe disease in man; the reported case-fatality rate is 3%–30%. The aim of this study was to determine the epidemiological and clinical characteristics of the CCHF cases in our clinic between 2009-2013. Materials and Methods: Thirty-three patients with the diagnosis of CCHF were followed up in Kayseri Training and Research Hospital between January 2009 and September 2013. Demographic, geographic, climatic, and clinical and laboratory characteristics of all patients were investigated. All of the cases were confirmed by CCHF immunoglobulin M (IgM) and/or PCR RNA positivity. Results: According to our reports 33 CCHF cases were followed in our clinic. Of the CCHF cases, 63.6% were male. Thirty patients (90.9%) were from rural regions. Seventeen patients (51, 5%) were farmers. The median age was 46.7 years (range18-71 years). On admission, 97% of patients experienced high fever, 100% had weakness, 93.9% had headache. The disease was more usual in May, June, July. Of the CCHF cases, 69.7% had a history of tick bite. On admission, all of the patients had thrombocytopenia, 87.9% had leucopenia, 27.3% had anemia, and 87.9% had elevated AST and ALT. Oral ribavirin treatment was used in 33.3% of the CCHF cases. The case-fatality rate was 12.1% (4/33 patients). Conclusions: CCHF remains a seasonal problem in the Mid-Eastern Anatolia region of Turkey. The mortality rate in our patients was higher than reported in other studies in our country (12% vs 5%). CCHF should be accompanied with supportive care, especially including early platelet replacement.

Severe Fever with Thrombocytopenia Syndrome Virus, Crimean-Congo Haemorrhagic Fever Virus, and Migratory Birds

Severe Fever with Thrombocytopenia Syndrome (SFTS) and Crimean-Congo Haemorrhagic Fever (CCHF) are tick-borne diseases belonging to the family Bunyaviridae. Since SFTS was first reported in China in 2009, the virus was isolated and confirmed in 2011, with additional reports of SFTSV expanding its geographic range from China to South Korea and Japan. CCHFV has the widest geographic distribution of any tick-borne virus, encompassing around 30 countries from eastern China through Asia, the Middle East, and southeastern Europe to Africa. During the past decade, CCHFV has emerged in new areas of Europe, Africa, the Middle East, and Asia and has increased in endemic areas. Migratory birds are considered to play a role in dispersing CCHFV vectors, and the virus. This review summarises SFTSV and CCHFV, highlighting the role of migratory birds in the transmission of tick-borne disease.

Does electrocardiography at admission predict outcome in Crimean-Congo hemorrhagic fever.

Background & objectives: Crimean-Congo hemorrhagic fever is an acute viral hemorrhagic fever with considerable mortality. Despite increasing knowledge about hemorrhagic fever viruses, the pathogenesis of Crimean-Congo hemorrhagic fever and causes of death were not well described. We aimed to evaluate whether there were electrocardiographic parameters designating mortality among these patients. Study design: This retrospective study was performed among confirmed Crimean-Congo hemorrhagic fever cases in Turkey. Electrocardiography was available in 49 patients within 24 h of hospitalization. All electrocardiograms were evaluated by two expert cardiologists according to Minnesota coding system. Results: Among patients with available electrocardiograms, there were 31 patients who survived, and 18 patients who died of Crimean-Congo hemorrhagic fever. Both groups were similar in terms of age, sex, body temperature, heart rate, and blood parameters. T-wave changes and bundle branch block were more frequently encountered among those who died. Presence of T-wave negativity or bundle branch block in this cohort of patients with Crimean-Congo hemorrhagic fever predicted death with a sensitivity of 72.7%, specificity of 92.6%, positive predictive value of 88.9%, negative predictive value of 80.6%. Conclusions: We think within the light of our findings that simple electrocardiography at admission may help risk stratification among Crimean-Congo hemorrhagic fever cases.

Epidemiology and Mutational Analysis of Global Strains of Crimean-Congo Haemorrhagic Fever Virus / 中国病毒学·英文版

Crimean-Congo hemorrhagic fever (CCHF) is a severe illness with high fatality.Cases are reported in several countries in Africa,Europe,the Middle East,and Asia.Phylogenetic analyses based on the virus S (nucleocapsid),M (glycoprotein),and L (polymerase) genome segments sequences indicate distinct geographic lineages exist but their specific genetic characteristics require elucidation.In this work we collected all full length S segment sequences and generated a phylogenetic tree based on the alignment of these 62 samples.We then analyzed the alignment using entries from AAIndex,the Amino Acid Index database,to identify amino acid mutations that performed significant changes in charge,pka,hydropathy and side chain volume.Finally,we mapped these changes back to the tree and alignment to identify correlated mutations or sites that characterized a specific lineage.Based on this analysis we are able to propose a number of sites that appear to be important for virus function and which would be good candidates for experimental mutational analysis studies.

Production of CCHF Virus-Like Particle by a Baculovirus-Insect Cell Expression System / 中国病毒学·英文版

Crimean-Congo Haemorrhagic Fever Virus(CCHFV)is a tick-born virus of the Nairovirus genus within the Bunyaviridae family,which is widespread and causes,high fatality. The nucleocapsid of CCHFV is comprised of N proteins that are encoded by the S segment. In this research,the N protein of CCHFV was expressed in insect cells using a recombinant baculovirus. Under an electron microscope,Virus-Like Particles (VLPs)with various size and morphology were observed in cytoplasmic vesicles in the infected cells.Sucrose-gradient purification of the cell lysate indicated that the VLPs were mainly located in the upper fraction after ultracentrifugation,which was confirmed by Western blot analysis and immuno-electron microscopy(IEM).

A new perspective to determine the severity of cases with Crimean-Congo hemorrhagic fever.

Background & objectives: We have established a severity grading score (SGS) system for predicting the fatality in Crimean-Congo hemorrhagic fever (CCHF) for the first time. Methods: This SGS has been set up by using several variables which were assumed to be associated with mortality according to the literature and also were considered to have clinical importance. Results: In all, 237 patients who had symptoms of CCHF for <5 days were included. The patients were grouped into three categories according to the mortality risk by using SGS as follows : low or no risk, intermediate and high risk groups. A SGS <5 showed no association with mortality (there were 158 cases in this group and all survived). This group constituted 66.7% of all the patients with CCHF. A SGS 6–10 showed moderate risk of mortality (10%) and seven out of 70 patients in this group died. SGS >11 means high risk for mortality (67%) and six out of 9 patients in this group died (p = 0.001). The sensitivity, specificity, accuracy, positive predictive value, and negative predictive value for >11 points of SGS were 67, 100, 98, 100, and 98%, respectively. Conclusions: This scoring system may help the clinicians to decide which patient to refer to a tertiary step hospital which may also decrease the cost and improve the functionality of healthcare staff.

Determination of serum adenosine deaminase and xanthine oxidase levels in patients with crimean-congo hemorrhagic fever

OBJECTIVE: Crimean-Congo hemorrhagic fever is an acute viral hemorrhagic fever with a high mortality rate. Despite increasing knowledge about hemorrhagic fever viruses, little is known about the pathogenesis of Crimean-Congo hemorrhagic fever. In this study, we measured serum adenosine deaminase and xanthine oxidase levels in Crimean-Congo hemorrhagic fever patients. METHODS: Serum adenosine deaminase levels were measured with a sensitive colorimetric method described by Giusti and xanthine oxidase levels by the method of Worthington in 30 consecutive hospitalized patients (mean age 42.6 ± 21.0). Laboratory tests confirmed their diagnoses of Crimean-Congo hemorrhagic fever. Thirty-five subjects (mean age 42.9 ± 19.1) served as the control group. RESULTS: There was a significant difference in adenosine deaminase and xanthine oxidase levels between cases and controls (p<0.05). However, neither adenosine deaminase nor xanthine oxidase levels varied with the severity of disease in the cases assessed (p>0.05). CONCLUSION: Adenosine deaminase and xanthine oxidase levels were increased in patients with Crimean-Congo hemorrhagic fever. Elevated serum xanthine oxidase activity in patients with Crimean-Congo hemorrhagic fever may be associated with reactive oxygen species generated by the xanthine/xanthine oxidase system during inflammatory responses. In addition, elevated lipid peroxidation may contribute to cell damage and hemorrhage. The association of cell damage and hemorrhage with xanthine oxidase activity should be further investigated in large-scale studies.

Serial Expression of the Truncated Fragments of the Nucleocapsid Protein of CCHFV and Identification of the Epitope Region / 中国病毒学·英文版

The Crimean-congo hemorrhagic fever virus(CCHFV)is a geographically widespread fatal pathogen.Identification of the epitope regions of the virus is important for the diagnosis and epidemiological studies of CCHFV infections.In this study,expression vectors carrying series truncated fragments of the NP(nueleoeapsid protein)gene from the S fragment of CCHFV strain YL04057 were constructed.The recombinant proteins were expressed in E.coli and purified for detection.The antigenic of the truncated fragments of NP was detected with a polyclonai serum(rabbit)and 2 monoclonal(mAbs)(14B7 and 43E5)against CCHFV by Western-blot analyses.The results showed that the three expressed constructs,which all contained the region 235AA to 305AA could be detected by mAbs polyclonal serum.The results suggest that region 235-305 aa of NP is a highly antigenic region and is highly conserved in the NP protein.