RESUMO
Zika virus (ZIKV) RNA has been found to remain in human semen for up to one year after infection, but the presence of Flavivirus antigens in the different compartments of semen has been largely unexplored. Following the introduction of ZIKV in Nicaragua (2016), a prospective study of patients with clinical symptoms consistent with ZIKV was conducted in León to investigate virus shedding in different fluids. ZIKV infection was confirmed in 16 male subjects (≥18 years of age) by RT-qPCR in either blood, saliva or urine. Of these, three provided semen samples at 7, 14, 21, 28, 60 and 180 days postsymptom onset (DPSO) for Flavivirus antigens and RNA studies. These cases were compared with 19 asymptomatic controls. Flavivirus antigens were examined by immunofluorescence (IF) using the 4G2 Mabs, and confocal microscopy was used to explore fluorescence patterns. The three (100%) symptomatic subjects and 3 (16%) of the 19 asymptomatic subjects had Flavivirus antigens and viral RNA in the spermatozoa fraction. The percentage of IF Flavivirus-positive spermatozoa cells ranged from 1.9% to 25% in specimens from symptomatic subjects, as compared with 0.8% to 3.8% in specimens from asymptomatic controls. A marked IF-pattern in the cytoplasmic droplets and tail of the spermatozoa was observed. The sperm concentrations (45 × 106/mL vs. 63.5 × 106/mL, p = 0.041) and the total motility percentage (54% vs. 75%, p = 0.009) were significantly lower in specimens from ZIKV-positive than in those of ZIKV-negative. In conclusion, this study demonstrated the presence of Flavivirus antigens and RNA within a time frame of 28 DPSO in sperm cells of symptomatic and asymptomatic subjects during the ZIKV epidemic. These findings have implications for public health, in terms of nonarthropod-born, silent transmission facilitated by sperm cells and potential transmission from asymptomatic males to pregnant women, with consequences to the fetus.
Assuntos
Antígenos Virais/análise , Flavivirus/isolamento & purificação , RNA Viral/análise , Espermatozoides/virologia , Infecção por Zika virus/virologia , Zika virus/isolamento & purificação , Adulto , Antígenos Virais/sangue , Antígenos Virais/urina , Flavivirus/genética , Imunofluorescência , Humanos , Masculino , RNA Viral/sangue , RNA Viral/urina , Reação em Cadeia da Polimerase em Tempo Real , Saliva/virologia , Sêmen/virologia , Espermatozoides/química , Eliminação de Partículas Virais , Adulto Jovem , Zika virus/genéticaRESUMO
Hantavirus cardiopulmonary syndrome (HCPS) is a highly pathogenic emerging disease (40% case fatality rate) caused by New World hantaviruses. Hantavirus infections are transmitted to humans mainly by inhalation of virus-contaminated aerosol particles of rodent excreta and secretions. At present, there are no antiviral drugs or immunotherapeutic agents available for the treatment of hantaviral infection, and the survival rates for infected patients hinge largely on early virus recognition and hospital admission and aggressive pulmonary and hemodynamic support. In this study, we show that Andes virus (ANDV) interacts with human apolipoprotein H (ApoH) and that ApoH-coated magnetic beads or ApoH-coated enzyme-linked immunosorbent assay plates can be used to capture and concentrate the virus from complex biological mixtures, such as serum and urine, allowing it to be detected by both immunological and molecular approaches. In addition, we report that ANDV-antigens and infectious virus are shed in urine of HCPS patients.
Assuntos
Antígenos Virais/urina , Síndrome Pulmonar por Hantavirus/urina , Orthohantavírus/imunologia , beta 2-Glicoproteína I/imunologia , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Antígenos Virais/sangue , Antígenos Virais/imunologia , Chlorocebus aethiops , Ensaio de Imunoadsorção Enzimática , Síndrome Pulmonar por Hantavirus/sangue , Síndrome Pulmonar por Hantavirus/imunologia , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Microesferas , RNA Viral/análise , Células VeroRESUMO
The dot ELISA technique was applied for direct detection of BK virus in clinical urine samples. The assay was performed on nitrocellulose paper dotted with the polyethylene glycol precipitated urine samples free of cellular debris. BK virus was detected with an anti-BK virus monoclonal antibody, and the complex was visualized by immunoperoxidase staining. Positive reaction appeared as well-defined dark blue spots. Of the 110 urine samples examined, 31 were positive in the dot ELISA and 79 proved negative. Comparing with the IIF results, the dot ELISA had a 88.46% of sensibility and 90.4% of specificity, and the results agreed completely in 99 samples. The simple dot ELISA technique can be recommended for detection of BK virus excretion in routinary diagnostic.
Assuntos
Antígenos Virais/urina , Vírus BK/imunologia , Infecções Tumorais por Vírus/diagnóstico , Animais , Anticorpos Monoclonais/imunologia , Precipitação Química , Ensaio de Imunoadsorção Enzimática , Humanos , Immunoblotting , Imunoglobulina G/imunologia , Peroxidase , Células VeroRESUMO
A procedure that allows to ascertain a diagnosis of Argentine Hemorrhagic Fever (AHF) as early as 24-48 hours after onset of symptoms is described. An immunofluorescent (IF) test on round cells of urinary sediment was employed. The procedure was assayed on 31 patients with febrile syndrome during epidemic peaks of 1975-1976. It was positive in 19 and negative in 12 cases. The 19 positive cases were confirmed AHF by clinical follow up and serology. From 12 negative cases, 8 belonged to other etiologies and 2 were confirmed AHF. The usefulness of the procedure for early diagnosis is emphasized.