RESUMEN
We determined levels of tick-borne encephalitis (TBE) virus (TBEV) RNA in serum samples obtained from 80 patients during the initial phase of TBE in Slovenia. For most samples, levels were within the range of 3-6 log10 copies RNA/mL. Levels were higher in female patients than in male patients, but we found no association between virus load and several laboratory and clinical parameters, including severity of TBE. However, a weak humoral immune response was associated with a more severe disease course, suggesting that inefficient clearance of virus results in a more serious illness. To determine whether a certain genetic lineage of TBEV had a higher virulence potential, we obtained 56 partial envelope protein gene sequences by directly sequencing reverse transcription PCR products from clinical samples of patients. This method provided a large set of patient-derived TBEV sequences. We observed no association between phylogenetic clades and virus load or disease severity.
Asunto(s)
Virus de la Encefalitis Transmitidos por Garrapatas , Encefalitis Transmitida por Garrapatas/epidemiología , Encefalitis Transmitida por Garrapatas/virología , ARN Viral , Carga Viral , Adulto , Anticuerpos Antivirales/inmunología , Virus de la Encefalitis Transmitidos por Garrapatas/genética , Virus de la Encefalitis Transmitidos por Garrapatas/inmunología , Femenino , Humanos , Inmunoglobulina G/inmunología , Masculino , Persona de Mediana Edad , Filogenia , Eslovenia/epidemiologíaRESUMEN
Crimean-Congo hemorrhagic fever (CCHF) is a highly contagious viral tick-borne disease with case-fatality rates as high as 50%. We describe a collaborative evaluation of the characteristics, performance, and on-site applicability of serologic and molecular assays for diagnosis of CCHF. We evaluated ELISA, immunofluorescence, quantitative reverse transcription PCR, and low-density macroarray assays for detection of CCHF virus using precharacterized archived patient serum samples. Compared with results of local, in-house methods, test sensitivities were 87.8%-93.9% for IgM serology, 80.4%-86.1% for IgG serology, and 79.6%-83.3% for genome detection. Specificity was excellent for all assays; molecular test results were influenced by patient country of origin. Our findings demonstrate that well-characterized, reliable tools are available for CCHF diagnosis and surveillance. The on-site use of such assays by health laboratories would greatly diminish the time, costs, and risks posed by the handling, packaging, and shipping of highly infectious biologic material.
Asunto(s)
Virus de la Fiebre Hemorrágica de Crimea-Congo/aislamiento & purificación , Fiebre Hemorrágica de Crimea/diagnóstico , Ensayo de Inmunoadsorción Enzimática , Virus de la Fiebre Hemorrágica de Crimea-Congo/genética , Virus de la Fiebre Hemorrágica de Crimea-Congo/inmunología , Humanos , Análisis por Micromatrices , Reacción en Cadena de la Polimerasa , Juego de Reactivos para Diagnóstico , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Pruebas SerológicasRESUMEN
BACKGROUND: Hantaviruses are the causative agents of two zoonotic diseases: hemorrhagic fever with renal syndrome (HFRS) and hantavirus cardiopulmonary syndrome (HCPS). The pathogenesis of HFRS is poorly understood. However, it has been suggested that immune mechanisms, including cytokines, might have an important role in HFRS pathogenesis. Thus, the aim of our study was to investigate cytokine profiles in serum samples of HFRS patients from Slovenia and explore a possible correlation between cytokine levels and disease severity. METHODS: Acute-phase serum samples from 52 patients, diagnosed with DOBV infection, and 61 patients, diagnosed with PUUV infection, were included in this study. Patients were divided into two groups--severe or mild--based on disease severity. Levels of IL-10, IL-12, INF-γ and TNF-α were measured in the serum samples with commercial ELISA tests. RESULTS: Increased levels of IL-10, INF-γ, and TNF-α were found in almost all the serum samples tested. On average, higher concentrations were detected in patients infected with DOBV than PUUV. Furthermore, significantly higher levels of IL-10 (P=0.001) and TNF-α (P=0.003) were found in patients with a more severe clinical course of disease. The same association between IL-10 (P<0.001) and TNF-α (P=0.021), and the severity of the disease was observed also when only patients infected with DOBV were considered. No differences in cytokine concentrations according to disease severity were observed in patients infected with PUUV. Concentrations of serum IL-12 in HFRS patients were in the normal range, however, higher levels were detected in patients infected with PUUV than in patients infected with DOBV. CONCLUSIONS: We suggest that imbalance in production of proinflammatory and regulatory cytokines might be in part responsible for a more severe course of HFRS.
Asunto(s)
Citocinas/sangre , Fiebre Hemorrágica con Síndrome Renal/inmunología , Mediadores de Inflamación/sangre , Adulto , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Citocinas/inmunología , Femenino , Orthohantavirus/inmunología , Fiebre Hemorrágica con Síndrome Renal/sangre , Fiebre Hemorrágica con Síndrome Renal/virología , Humanos , Mediadores de Inflamación/inmunología , MasculinoRESUMEN
In hantavirus infections levels of serum leukocytes or C-reactive protein are usually elevated to levels found in serious bacterial infections. However, procalcitonin in patients infected with hantavirus has not yet been discussed in the literature. A total of 29 adult patients with hantavirus infection, 30 with sepsis, and 19 with tick-borne encephalitis were included in this observational retrospective study. The median procalcitonin level in patients with hantavirus infection was 0.53 µg/L (range 0.09-11.71 µg/L), in the group with sepsis 4.33 µg/L (range 0.08-161.1 µg/L) and in patients with viral meningitis 0.08 µg/L (range 0.05-0.12 µg/L). The difference between all three groups was statistically significant (p < 0.001). A higher procalcitonin level was found in patients with hemorrhagic fever with renal syndrome caused by Dobrava virus (0.74 µg/L; range 0.09-2.83 µg/L) than in those with Puumala virus infections (0.50 µg/L; range 0.10-11.71 µg/L). However, the difference was not statistically significant (p = 0.895). This study confirmed previous findings demonstrating the association of elevated procalcitonin with bacterial infection. However, increased procalcitonin serum level was also found in hantavirus infections with overlapping results between viral and severe bacterial infections.
Asunto(s)
Calcitonina/sangre , Infecciones por Hantavirus/sangre , Precursores de Proteínas/sangre , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Bacteriemia/sangre , Bacteriemia/microbiología , Bacteriemia/mortalidad , Péptido Relacionado con Gen de Calcitonina , Niño , Encefalitis Transmitida por Garrapatas/sangre , Encefalitis Transmitida por Garrapatas/mortalidad , Femenino , Humanos , Masculino , Meningitis Viral/sangre , Persona de Mediana Edad , Estudios Retrospectivos , Sepsis/sangre , Sepsis/microbiología , Sepsis/mortalidad , Adulto JovenRESUMEN
Hemorrhagic fever with renal syndrome (HFRS) and Crimean-Congo hemorrhagic fever (CCHF) are important viral hemorrhagic fevers (VHF), especially in the Balkan region. Infections with Dobrava or Puumala orthohantavirus and Crimean-Congo hemorrhagic fever orthonairovirus can vary from a mild, nonspecific febrile illness, to a severe disease with a fatal outcome. The pathogenesis of both diseases is poorly understood, but it has been suggested that a host's immune mechanism might influence the pathogenesis of the diseases and survival. The aim of our study is to characterize cytokine response in patients with VHF in association with the disease progression and viral load. Forty soluble mediators of the immune response, coagulation, and endothelial dysfunction were measured in acute serum samples in 100 HFRS patients and 70 CCHF patients. HFRS and CCHF patients had significantly increased levels of IL-6, IL-12p70, IP-10, INF-γ, TNF-α, GM-CSF, MCP-3, and MIP-1b in comparison to the control group. Interestingly, HFRS patients had higher concentrations of serum MIP-1α, MIP-1ß, which promote activation of macrophages and NK cells. HFRS patients had increased concentrations of IFN-γ and TNF-α, while CCHF patients had significantly higher concentrations of IFN-α and IL-8. In both, CCHF and HFRS patients' viral load significantly correlated with IP-10. Patients with fatal outcome had significantly elevated concentrations of IL-6, IFN-α2 and MIP-1α, while GRO-α, chemokine related to activation of neutrophils and basophils, was downregulated. Our study provided a comprehensive characterization of biomarkers released in the acute stages of CCHF and HFRS.
Asunto(s)
Citocinas/sangre , Fiebre Hemorrágica con Síndrome Renal/inmunología , Fiebre Hemorrágica de Crimea/inmunología , Anticuerpos Antivirales/sangre , Biomarcadores/sangre , Quimiocina CXCL10/sangre , Progresión de la Enfermedad , Femenino , Orthohantavirus/inmunología , Virus de la Fiebre Hemorrágica de Crimea-Congo/inmunología , Fiebre Hemorrágica con Síndrome Renal/patología , Fiebre Hemorrágica con Síndrome Renal/virología , Fiebre Hemorrágica de Crimea/patología , Fiebre Hemorrágica de Crimea/virología , Humanos , Masculino , Eslovenia , Carga ViralRESUMEN
The Balkan region and Kosovo in particular, is a well-known Crimean-Congo hemorrhagic fever (CCHF) endemic region, with frequent epidemic outbreaks and sporadic cases occurring with a hospitalized case fatality of approximately 30%. Recent analysis of complete genome sequences of diverse CCHF virus strains showed that the genome plasticity of the virus is surprisingly high for an arthropod-borne virus. High levels of nucleotide and amino acid differences, frequent RNA segment reassortment and even RNA recombination have been recently described. This diversity illustrates the need to determine the complete genome sequence of CCHF virus representatives of all geographically distinct endemic areas, particularly in light of the high pathogenicity of the virus and its listing as a potential bioterrorism threat. Here we describe the first complete CCHF virus genome sequence of a virus (strain Kosova Hoti) isolated from a hemorrhagic fever case in the Balkans. This virus strain was isolated from a fatal CCHF case, and passaged only twice on Vero E6 cells prior to sequence analysis. The virus total genome was found to be 19.2 kb in length, consisting of a 1672 nucleotide (nt) S segment, a 5364 nt M segment and a 12150 nt L segment. Phylogenetic analysis of CCHF virus complete genomes placed the Kosova Hoti strain in the Europe/Turkey group, with highest similarity seen with Russian isolates. The virus M segments are the most diverse with up to 31 and 27% differences seen at the nt and amino acid levels, and even 1.9% amino acid difference found between the Kosova Hoti and another strain from Kosovo (9553-01). This suggests that distinct virus strains can coexist in highly endemic areas.
Asunto(s)
Brotes de Enfermedades , Genoma Viral , Virus de la Fiebre Hemorrágica de Crimea-Congo/genética , Fiebre Hemorrágica de Crimea/epidemiología , Fiebre Hemorrágica de Crimea/virología , Animales , Chlorocebus aethiops , Europa (Continente) , Femenino , Virus de la Fiebre Hemorrágica de Crimea-Congo/clasificación , Humanos , Filogenia , Recombinación Genética , Homología de Secuencia , Pase Seriado , Células Vero , Yugoslavia/epidemiologíaRESUMEN
Early and accurate diagnosis of Crimean-Congo hemorrhagic fever (CCHF) is essential for the treatment and outcome of the disease and prevention of its further transmission. Molecular-based diagnostic assays now serve as the front-line tool in the diagnosis of CCHF. However, the development of real-time RT-PCR assay for the detection of Crimean-Congo hemorrhagic fever virus (CCHFV) has been hampered by a virus strain variation. The development of a one-step real-time RT-PCR assay for the detection of CCHFV is described herein. The technique is based on the fluorescence resonance energy transfer probe technology employing the endonuclease activity of Taq polymerase enzyme. The assay was designed to detect specifically the strains from a phylogenetic cluster of CCHFV which encompasses the known CCHFV strains circulating in the Balkan region. The detection system was tested using CCHFV strain Kosovo Hoti, clinical serum samples and ticks. The real-time assay described is rapid, specific and sensitive. Since the Balkan peninsula is also an endemic region for hemorrhagic fever with renal syndrome (HFRS), this method is suggested as convenient for early differential diagnosis of suspected viral hemorrhagic fever patients.
Asunto(s)
Virus de la Fiebre Hemorrágica de Crimea-Congo/genética , Virus de la Fiebre Hemorrágica de Crimea-Congo/aislamiento & purificación , Fiebre Hemorrágica de Crimea/diagnóstico , Fiebre Hemorrágica de Crimea/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Animales , Benzotiazoles , Chlorocebus aethiops , Diaminas , Estudios de Evaluación como Asunto , Colorantes Fluorescentes , Fiebre Hemorrágica de Crimea/sangre , Humanos , Compuestos Orgánicos , Quinolinas , ARN Viral/sangre , Juego de Reactivos para Diagnóstico , Sensibilidad y Especificidad , Polimerasa Taq , Garrapatas/virología , Factores de Tiempo , Células Vero , Yugoslavia/epidemiologíaRESUMEN
BACKGROUND: Tick-borne encephalitis virus (TBEV) is one of the most important causes of human viral infections of the central nervous system in Europe. Currently, the diagnosis of TBE is based on the demonstration of specific antibodies in patient's serum, which appear only several weeks after the infection. OBJECTIVE: To determine how successfully can viral RNA be detected by RT-PCR in the samples of body fluids of patients with TBE prior to and after the appearance of antibodies. STUDY DESIGN: Serum, whole blood and CSF samples from 34 patients with a serologically confirmed TBE were collected. Samples were tested for the presence of TBEV RNA by using RT-PCR method. RESULTS: Viral RNA was detected in all blood and serum samples collected before the development of antibodies. After the appearance of IgM antibodies, the number of positive samples dropped by at least one third. After the development of IgG antibodies, only 3% of serum and 16% of blood samples tested positive for viral RNA. Samples of cerebrospinal fluid were shown to be inappropriate for the molecular diagnosis of TBE using this assay, since only one sample (10%) that was collected in the sero-negative phase of disease was found positive by the PCR assay. CONCLUSIONS: RT-PCR is an efficient method for an early detection of TBEV in blood and serum samples collected prior to the appearance of antibodies. This method can be of valuable use for a differential diagnosis of TBEV infection in patients with febrile illness after a tick bite, particularly in regions where more than one tick-transmitted diseases are endemic.
Asunto(s)
Virus de la Encefalitis Transmitidos por Garrapatas/aislamiento & purificación , Encefalitis Transmitida por Garrapatas/diagnóstico , ARN Viral/sangre , ARN Viral/líquido cefalorraquídeo , Adolescente , Adulto , Animales , Anticuerpos Antivirales/sangre , Encéfalo/virología , Diagnóstico Diferencial , Virus de la Encefalitis Transmitidos por Garrapatas/clasificación , Virus de la Encefalitis Transmitidos por Garrapatas/genética , Virus de la Encefalitis Transmitidos por Garrapatas/inmunología , Encefalitis Transmitida por Garrapatas/microbiología , Humanos , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Crimean-Congo hemorrhagic fever (CCHF) is an acute, tick borne disease often associated with hemorrhagic presentations and high case fatality rate. Kosovo is a highly endemic area for CCHF, with a significant case fatality rate. The aim of our study was to determine the prevalence of CCHF in Kosovo. We tested 1105 serum samples from healthy population in both endemic and non-endemic areas in the country. Our results revealed a seroprevalence of 4.0% (range 0-9.3%) which is comparable to the seroprevalence in other countries. We show that seroprevalence is correlated to the disease incidence in each studied municipality. We also tested 401 animal sera (353 cow, 30 sheep, 10 goat and 8 chicken) in four endemic municipalities in Kosovo. We detected specific antibodies in all animals except in chicken. Seroprevalence in cows is comparable to other endemic areas and correlates to the seroprevalence in humans. No CCHF RNA could be detected in 105 tick samples obtained in 2012 and 2013. Sequencing of CCHFV positive ticks from 2001 revealed that the virus is most closely related to viral strains that were detected in CCHF patients from Kosovo. Results suggest that mild CCHF cases are most probably underdiagnosed and consequently that the burden of disease is higher than reported. Our study provides key information for CCHF surveillance and raises awareness for possible imported cases in CCHF non-endemic countries.
Asunto(s)
Anticuerpos Antivirales/sangre , Virus de la Fiebre Hemorrágica de Crimea-Congo/inmunología , Fiebre Hemorrágica de Crimea/epidemiología , Fiebre Hemorrágica de Crimea/veterinaria , Ganado/virología , Adulto , Anciano , Animales , Anticuerpos Antivirales/inmunología , Secuencia de Bases , Bovinos , Pollos/virología , ADN/genética , Femenino , Cabras/virología , Virus de la Fiebre Hemorrágica de Crimea-Congo/clasificación , Virus de la Fiebre Hemorrágica de Crimea-Congo/genética , Fiebre Hemorrágica de Crimea/virología , Humanos , Inmunoglobulina G/sangre , Kosovo/epidemiología , Ganado/inmunología , Masculino , Persona de Mediana Edad , Enfermedades de las Aves de Corral/virología , Prevalencia , Análisis de Secuencia de ADN , Estudios Seroepidemiológicos , Ovinos/virología , Enfermedades por Picaduras de Garrapatas/transmisión , Garrapatas/virología , Adulto JovenRESUMEN
BACKGROUND: Phleboviruses are large and widespread group of viruses that are transmitted by arthropods and they have been reported to circulate in endemic regions of Mediterranean Basin, including Croatia. OBJECTIVES: To investigate the role of Toscana virus, as a cause of the aseptic meningitis, in summer months in Croatia. STUDY DESIGN: Samples from 30 patients with aseptic meningitis were retrospectively tested by serology and RT-PCR for TOSV. RESULTS: TOSV RNA was detected in 2/30 and TOSV IgM antibodies were found in 4/30 of patients. Phylogenetic analysis of partial L and S segments suggests that TOSV from Croatia represents an autochthonous strain. CONCLUSIONS: The study has confirmed the role of TOSV as an agent that causes aseptic meningitis in Croatia, therefore it should be considered by physicians when encountering meningitis or febrile illness among indigenous population or travellers during the summer months.
Asunto(s)
Infecciones por Bunyaviridae/virología , Meningitis Aséptica/virología , Virus de Nápoles de la Fiebre de la Mosca de los Arenales/aislamiento & purificación , Adolescente , Adulto , Anticuerpos Antivirales/sangre , Infecciones por Bunyaviridae/sangre , Infecciones por Bunyaviridae/diagnóstico , Niño , Preescolar , Croacia , Femenino , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Masculino , Meningitis Aséptica/sangre , Meningitis Aséptica/diagnóstico , Persona de Mediana Edad , Filogenia , ARN Viral/sangre , ARN Viral/líquido cefalorraquídeo , Estudios Retrospectivos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Virus de Nápoles de la Fiebre de la Mosca de los Arenales/genéticaRESUMEN
The hard tick Ixodes ricinus is the principal vector of tick-borne encephalitis virus (TBEV) in Slovenia; but until now, there was no information about the prevalence of TBEV infection in Slovenian ticks. We conducted a 2-year survey in 2005 and 2006, during which we were collecting I. ricinus ticks monthly in eight different locations of Slovenia. A total of 4777 I. ricinus ticks were collected: 1515 in year 2005 and 3262 in year 2006. The collected ticks were pooled into groups from which total RNA was extracted. Viral RNA was detected using real-time RT-polymerase chain reaction (PCR). Ticks infected with TBEV were found in six of eight locations. Viral RNA was detected in 8 of the 230 pools of ticks collected in 2005 and in 14 of the 442 pools collected in 2006. Prevalence of TBEV infection in Slovenian ticks was determined as 0.47%: 0.54% in 2005 and 0.43% in 2006. The detected infection rate in ticks significantly correlates with the TBEV incidence rates in selected areas. Using the method of sequencing, we have confirmed that the TBEV in ticks is genetically related to the TBEV in Slovenian patients.
Asunto(s)
Virus de la Encefalitis Transmitidos por Garrapatas/genética , Virus de la Encefalitis Transmitidos por Garrapatas/aislamiento & purificación , Ixodes/virología , Animales , Masculino , Ninfa/virología , Filogenia , ARN Viral/genética , ARN Viral/aislamiento & purificación , Eslovenia , Factores de TiempoRESUMEN
Major histocompatibility complex (MHC) class I and class II genes regulate the balance between appropriate aggressive responses and invading pathogens while minimizing the destruction of host tissue. Several studies have shown that in hemorrhagic fever with renal syndrome (HFRS) patients, the disease outcome is determined by a complex interaction between the virus and immunopathologic and human genetic factors. In Slovenia, the severity of the disease caused by Puumala virus (PUUV) is significantly lower than that of HFRS due to Dobrava virus (DOBV). We have determined 23 different HLA-B and 12 different HLA-DRB1 types in Slovenian HFRS patients. Comparison of HLA frequencies between healthy individuals and HFRS patients showed no strong association with the susceptibility for hantaviral infection. Significant associations were recognized when the patient group was separated according to the virus responsible for the infection. DOBV-infected patients have a significantly higher frequency of HLA-B*35 than PUUV-infected patients. For HLA class II genes, the biggest difference between the PUUV- and DOBV-infected groups of patients was in HLA-DRB1*13, where this phenotype was more frequent in PUUV-infected patients, especially in the severe form of the disease. HLA-B*07 could play a protective role in PUUV-caused HFRS in the Slovenian population. Our study shows diverse associations of HLA molecules with DOBV- and PUUV-induced HFRS, and therefore, we presume that different hantaviruses are presented differently through the same HLA molecules and that this might lead to either a more severe or a milder form of the disease. In line with this idea, we have noticed that HLA-B*35 might be a genetic risk factor for DOBV infection in the Slovenian population.
Asunto(s)
Antígenos HLA-B/genética , Cadenas HLA-DRB1/genética , Fiebre Hemorrágica con Síndrome Renal/genética , Fiebre Hemorrágica con Síndrome Renal/fisiopatología , Orthohantavirus/patogenicidad , Virus Puumala/patogenicidad , Anticuerpos Antivirales/sangre , Progresión de la Enfermedad , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Antígenos HLA-B/clasificación , Antígenos HLA-B/inmunología , Cadenas HLA-DRB1/clasificación , Cadenas HLA-DRB1/inmunología , Orthohantavirus/inmunología , Fiebre Hemorrágica con Síndrome Renal/inmunología , Fiebre Hemorrágica con Síndrome Renal/virología , Prueba de Histocompatibilidad , Humanos , Inmunoglobulina M/sangre , Masculino , Virus Puumala/inmunología , Índice de Severidad de la Enfermedad , Eslovenia , Especificidad de la EspecieRESUMEN
Until now, the pathogenesis of Crimean-Congo hemorrhagic fever (CCHF) has not been well described. However, it has been hypothesized that it could be a result of the direct injury of virus-infected tissues in combination with the indirect effects of host immune responses, including cytokines. To shed more light on the role of viral load and cytokines, differential influences of CCHF virus (CCHFV) RNA load, antibody response, and cytokine production on severity and outcome of the disease were studied in sera of 46 patients with confirmed acute CCHF from Kosovo. In this study, viral load proved to be strongly related to the severity and outcome of the disease, with higher viral loads detected in patients with fatal outcomes than in surviving patients. Also, patients with fatal outcome had on average a weaker antibody response, if one was present at all. High levels of interleukin-10 (IL-10), gamma interferon (IFN-gamma), and tumor necrosis factor alpha (TNF-alpha) were associated with poor outcome, since detected concentrations were highest in patients with fatal outcome and lowest in patients with moderate disease course. Additionally, a positive linear dependence between viral load and these cytokines was observed. Interestingly, reduced levels of IL-12 were detected in all CCHF patients. Our study favors the hypothesis that CCHF could be a result of a delayed and downregulated immune response caused by IL-10, which leads to an increased replication and spread of CCHFV throughout the body. This consequently triggers increased production of IFN-gamma and TNF-alpha, cytokines mediating vascular dysfunction, disseminated intravascular coagulation, organ failure, and shock.
Asunto(s)
Fiebre Hemorrágica de Crimea/etiología , Inmunidad , Carga Viral , Anticuerpos Antivirales/sangre , Citocinas/sangre , Fiebre Hemorrágica de Crimea/inmunología , Humanos , Interleucina-10/sangre , Pronóstico , ARN Viral/sangre , Estudios Retrospectivos , Índice de Severidad de la EnfermedadRESUMEN
Hantaviruses cause a lifelong and asymptomatic infection in naturally infected hosts as well as in experimentally infected rodents. Understanding the ecology and pathogenesis of hantaviruses requires an interdisciplinary research approach, which links laboratory experiments with results gained from field studies. Although several studies report hantavirus persistence and tissue infection patterns for experimentally infected rodents, field data is very limited. For this reason, the aim of our study was to investigate Puumala, Dobrava and Saaremaa virus RNA loads and tissue infection patterns in their natural reservoirs. Hantavirus RNA was demonstrated in all tested internal organs and blood samples of 14 naturally infected rodent hosts. However, the concentration of a specific virus differs depending on the virus, the host and the organ tested. Above all, the Dobrava virus showed a considerably higher viral load in all internal organs and blood samples of infected Apodemus flavicollis hosts. Results obtained in the study support the thesis that virus RNA load reaches its peak in the first month after infection, presumably after the virus has spread throughout all internal organs. This also implies that recently infected rodents are more important for transmission of the virus in the community.
Asunto(s)
Infecciones por Hantavirus/veterinaria , Orthohantavirus/aislamiento & purificación , Virus Puumala/aislamiento & purificación , Roedores/virología , Carga Viral , Estructuras Animales/virología , Animales , Sangre/virología , Infecciones por Hantavirus/virología , ARN Viral/aislamiento & purificaciónRESUMEN
Ixodes ricinus is a vector of pathogens that cause many diseases in Europe and Slovenia: tick-borne encephalitis (TBE), anaplasmosis, borreliosis, babesiosis and others. The risk for contracting these diseases depends strongly on the density of the infected questing ticks and many studies have investigated tick population dynamics and the parameters affecting them. They have shown a clear influence of climatic and landscape arrangements in the microhabitat on tick abundance and dynamics and therefore on transmission of pathogens important in human and veterinary medicine. In our study we assessed the influence of climatic factors on questing activity of ticks over a three-year period at 7 locations in Slovenia. Locations were selected in endemic foci of TBE with different intensity, which were identified according to the presence of human disease. Sites differ according to various abiotic and biotic factors, such as climate, amount of rain, height above sea level, vegetation and wildlife. All three stages of ticks were collected monthly over a three-year period (2005-2007). Temperature, humidity and precipitation data were collected for these years. The purpose of our study was to relate observed differences in I. ricinus ticks questing activity to local climate. We found a correlation between the decrease of questing ticks in the summer and the combination of air temperatures and humidity in the form of saturation deficit.
Asunto(s)
Conducta Alimentaria/fisiología , Ixodes/fisiología , Animales , Clima , Calentamiento Global , Larva , Ninfa , Estaciones del Año , Eslovenia , Factores de TiempoRESUMEN
To asses the role of virus load in the pathogenesis of hemorrhagic fever with renal syndrome, the serum Dobrava virus RNA load in 46 patients was measured with a novel quantitative real-time reverse-transcriptase polymerase chain reaction assay and compared to the disease severity. The level of viremia, detected in 26 patients, ranged from 10(2)-10(8) copies/mL of serum. The patients with severe disease had, on average, higher viral RNA loads than patients with a milder course of disease (6.15 vs. 4.67 log(10) copies/mL; P = .053). These results suggest that the Dobrava virus load might be associated with the severity of disease.
Asunto(s)
Fiebre Hemorrágica con Síndrome Renal/sangre , Orthohantavirus/patogenicidad , ARN Viral/sangre , Carga Viral , Viremia/fisiopatología , Estudios de Cohortes , Progresión de la Enfermedad , Fiebre Hemorrágica con Síndrome Renal/fisiopatología , HumanosRESUMEN
We used quantitative real-time reverse transcription-PCR to measure viral load in serum from 24 patients in Kosovo who had acute Crimean-Congo hemorrhagic fever. Viral load correlated with clinical disease and antibodies and could be used as a predictor of disease outcome.