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1.
BMC Ophthalmol ; 23(1): 447, 2023 Nov 06.
Artículo en Inglés | MEDLINE | ID: mdl-37932668

RESUMEN

Uveitis is a process of intraocular inflammation that may involve different sections of the uveal tract. Apart from systemic or localized immune-mediated diseases, infections are key players in the etiology of uveitis and entail different treatment strategies. Rubella virus (RuV) is a recognized causative agent for the development of Fuchs uveitis, representing a major cause of virus-associated intraocular inflammation. A cohort of 159 patients diagnosed with different forms of uveitis between 2013 and 2019 was subjected to diagnostic antibody testing of the aqueous or vitreous humor. The diagnostic panel included RuV, cytomegalovirus, herpes simplex virus, varicella-zoster virus, and toxoplasmosis. Within this cohort, 38 RuV-associated uveitis (RAU) patients were identified based on a pathologic Goldman-Witmer coefficient indicative of an underlying RuV infection. With a mean age of 45.9 years, the RAU patients were younger than the non-RAU patients (56.3, p < 0.001). The evaluation of clinical parameters revealed a predominance of anterior uveitis and late sequalae such as cataract and glaucoma among the RAU patients. In 15 of the patients a history of prior RuV infections could be confirmed. The study underlines the importance of long-term surveillance of RuV associated diseases that originate from infections before the introduction of RuV vaccination programs.


Asunto(s)
Infecciones Virales del Ojo , Rubéola (Sarampión Alemán) , Enfermedades de la Úvea , Uveítis Anterior , Uveítis , Humanos , Persona de Mediana Edad , Virus de la Rubéola , Centros de Atención Terciaria , Infecciones Virales del Ojo/diagnóstico , Humor Acuoso , Rubéola (Sarampión Alemán)/diagnóstico , Uveítis Anterior/diagnóstico , Inflamación
2.
Virol J ; 19(1): 53, 2022 03 24.
Artículo en Inglés | MEDLINE | ID: mdl-35331278

RESUMEN

BACKGROUND: Hepatitis B virus (HBV) infection is a particular concern in human immunodeficiency virus (HIV) infected individuals. In Ethiopia, detailed clinical and virological descriptions of HBV prevailing during HIV co-infection and symptomatic liver disease patients are lacking. The aim of this study was to investigate HBV virological characteristics from Ethiopian HBV/HIV co-infected and HBV mono-infected individuals. METHODS: A total of 4105 sera from HIV positive individuals, liver disease patients, and blood donors were screened serologically for HBV. The overlapping polymerase/surface genome region of HBV from 180 infected individuals was extracted, amplified, and sequenced for genotypic analysis. RESULTS: The HBsAg seroprevalence was detected 43% in liver disease patients, 8.4% in blood donors, and 6.7% in HIV/HBV co-infected individuals. The occult HBV prevalence was 3.7% in HIV/HBV co-infected individuals and 2.8% in blood donors with an overall prevalence rate of 3.4%. A phylogenetic analysis showed three HBV genotypes; A (61.1%), D (38.3%) and E (0.6%). Genotype A belongs to subtypes A1 (99.1%) and A9 (0.9%), but genotype D showed heterogeneous subtypes; D2 (63.8%) followed by D4 (21.7%), D1 (8.7%), D3 (4.3%), and D10 (1.4%). CONCLUSIONS: The HIV/HBV co-infected individuals and blood donors showed lower HBsAg seroprevalence compared to liver diseases patients. Occult HBV prevalence showed no difference between HIV/HBV co-infected and blood donor groups. This study demonstrated predominance distribution of HBV subtypes A1 and D2 in northwest Ethiopia. The observed virological characteristics could contribute for evidence-based management of viral hepatitis in Ethiopia where antiretroviral therapy guidelines do not cater for viral hepatitis screening during HIV co-infection.


Asunto(s)
Coinfección , Infecciones por VIH , Hepatitis B , Coinfección/epidemiología , ADN Viral/genética , Etiopía/epidemiología , Infecciones por VIH/complicaciones , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/epidemiología , Hepatitis B/complicaciones , Hepatitis B/epidemiología , Antígenos de Superficie de la Hepatitis B , Virus de la Hepatitis B/genética , Humanos , Epidemiología Molecular , Mutación , Filogenia , Estudios Seroepidemiológicos
3.
Arch Virol ; 167(6): 1421-1431, 2022 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-35415782

RESUMEN

Human noroviruses (hNoVs) are an important cause of acute gastroenteritis worldwide. However, the lack of a reproducible in vitro cell culture system has impaired research and the development of preventive measures, therapeutic drugs, and vaccines. The aim of this study was to analyze and optimize a suitable cell line for in vitro cultivation of hNoV. The Caco-2 cell line, which is of colorectal origin and differentiates spontaneously into intestinal enterocyte-like cells, was chosen as a model. It was found that differentiated cells were more susceptible to infection with hNoV, resulting in a higher virus yield. This was accompanied by an increase in H type 1 antigen in the cell membrane during differentiation, which functions as an attachment factor for hNoV. Induced overexpression of H type 1 antigen in undifferentiated Caco-2 cells resulted in an increase in viral output to a level similar to that in differentiated cells. However, the relatively low level of viral output, which contrasts with what is observed in vivo, shows that the viral replication cycle is restricted in this model. The results indicate that there is a block at the level of viral release.


Asunto(s)
Infecciones por Caliciviridae , Gastroenteritis , Norovirus , Células CACO-2 , Técnicas de Cultivo de Célula , Humanos , Intestinos , Norovirus/genética
4.
Virol J ; 14(1): 176, 2017 09 12.
Artículo en Inglés | MEDLINE | ID: mdl-28899424

RESUMEN

BACKGROUND: Although hepatitis B virus (HBV) is hyperendemic and heterogeneous in its genetic diversity in Ethiopia, little is known about hepatitis D virus (HDV) circulating genotypes and molecular diversity. METHODS: A total of 321 hepatitis B surface antigen (HBsAg) positives (125 HIV co-infected, 102 liver disease patients and 94 blood donors) were screened for anti-HDV antibody. The anti-HDV positive sera were subjected to Real time PCR for HDV-RNA confirmation. The non coding genome region (spanning from 467 to 834 nucleotides) commonly used for HDV genotyping as well as complete HDV genome were sequenced for genotyping and molecular analysis. RESULTS: The anti-HDV antibody was found to be 3.2% (3) in blood donors, 8.0% (10) in HIV co-infected individuals and 12.7% (13) in liver disease patients. None of the HIV co-infected patients who revealed HBV lamivudine (3TC) resistance at tyrosine-methionine/isoleucine-aspartate-aspartate (YM(I)DD) reverse transcriptase (RT) motif with concomitant vaccine escape gene mutants was positive for anti-HDV antibody. The HDV viremia rate was 33.3%, 30.0% and 23.1% in respect to the above study groups. All the six isolates sequenced were phylogenetically classified as HDV genotype 1 (HDV-1) and grouped into two monophyletic clusters. Amino acid (aa) residues analysis of clathrin heavy chain (CHC) domain and the isoprenylation signal site (Py) at 19 carboxyl (C)-terminal amino acids (aa 196-214) and the HDV RNA binding domain (aa 79-107) were highly conserved and showed a very little nucleotide variations. All the sequenced isolates showed serine at amino acid position 202. The RNA editing targets of the anti-genomic HDV RNA (nt1012) and its corresponding genomic RNA (nt 580) showed nucleotides A and C, respectively. CONCLUSIONS: The low seroprevalence and viraemic rates of HDV in particular during HIV-confection might be highly affected by HBV drug resistance selected HBsAg mutant variants in this setting, although HDV-1 sequences analysis revealed clade homogeneity and highly conserved structural and functional domains. Thus, the potential role of HBV drug resistance associated polymerase mutations and concomitant HBsAg protein variability on HDV viral assembly, secretion and infectivity needs further investigation.


Asunto(s)
Variación Genética , Hepatitis D/epidemiología , Hepatitis D/virología , Virus de la Hepatitis Delta/clasificación , Virus de la Hepatitis Delta/genética , Filogenia , Adulto , Coinfección , Estudios Transversales , Etiopía/epidemiología , Femenino , Genotipo , Infecciones por VIH/complicaciones , Anticuerpos Antihepatitis/sangre , Hepatitis B/complicaciones , Hepatitis B/epidemiología , Hepatitis D/complicaciones , Hepatitis D/diagnóstico , Humanos , Masculino , Persona de Mediana Edad , Epidemiología Molecular , Análisis de Secuencia de ADN , Análisis de Secuencia de Proteína
6.
BMC Infect Dis ; 16(1): 761, 2016 12 19.
Artículo en Inglés | MEDLINE | ID: mdl-27993129

RESUMEN

BACKGROUND: The existing seroepidemiological data on viral hepatitis in Ethiopia showed a wide variation in prevalence pattern and the clinical and public health burden have been underestimated. The aim of this systematic review and meta-analysis was to provide a clear and comprehensive estimation of viral hepatitis epidemiology and the potential clinical burdens in Ethiopia. METHODS: A comprehensive literature search was carried out from five decades (1968-2015) published studies from biomedical databases; PubMed, Google scholar, Medline and Web of Science. RESULTS: The overall pooled prevalence of hepatitis B virus (HBV) was 7.4% (95%CI: 6.5-8.4). The pooled prevalence among subgroups showed 5.2% (95%CI: 3.7-7.4) in human immunodeficiency virus (HIV) infected individuals, 8.0% (95%CI: 5.9-10.7) in community based studies, 8.4% (95%CI: 5.4-12.7) in blood donors, 11.0% (95%CI: 7.5-15.9) in immigrants and 6.9% (95%CI: 5.6-8.5) in other groups. Among study parameters considered during meta-regression analysis, only study years were associated with a decreasing HBV prevalence rate over time. The overall pooled prevalence of anti-hepatitis C virus antibody (anti-HCV) was 3.1% (95%CI: 2.2-4.4). Unlike HBV, the anti-HCV prevalence in HIV infected individuals was higher (5.5%, 95%CI: 3.8-7.8%, p = 0.01) than the prevalence observed in the other subgroup of study population. Although relatively few data were available, hepatitis virus A (HAV), D (HDV) and E (HEV) were also circulated in Ethiopia. CONCLUSIONS: This review indicates that all types of viral hepatitis origins are endemic in Ethiopia. Adapting a recommended diagnostic and treatment algorithm of viral hepatitis in the routine healthcare systems and implementing prevention and control policies in the general population needs an urgent attention.


Asunto(s)
Hepatitis Viral Humana/epidemiología , Infecciones Oportunistas Relacionadas con el SIDA/epidemiología , Adolescente , Adulto , Donantes de Sangre/estadística & datos numéricos , Coinfección/epidemiología , Etiopía/epidemiología , Infecciones por VIH/epidemiología , Infecciones por VIH/virología , Hepatitis B/epidemiología , Hepatitis B/virología , Virus de la Hepatitis B/inmunología , Virus de la Hepatitis B/patogenicidad , Hepatitis C/epidemiología , Anticuerpos contra la Hepatitis C/sangre , Virus de Hepatitis/patogenicidad , Hepatitis Viral Humana/virología , Humanos , Persona de Mediana Edad , Adulto Joven
7.
J Transl Med ; 13: 377, 2015 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-26626277

RESUMEN

BACKGROUND: Although biochemical analysis of HIV-1 integrase enzyme suggested the use of integrase inhibitors (INIs) against HIV-1C, different viral subtypes may favor different mutational pathways potentially leading to varying levels of drug resistance. Thus, the aim of this study was to search for the occurrence and natural evolution of integrase polymorphisms and/or resistance mutations in HIV-1C Ethiopian clinical isolates prior to the introduction of INIs. METHODS: Plasma samples from chronically infected drug naïve patients (N = 45), of whom the PR and RT sequence was determined previously, were used to generate population based sequences of HIV-1 integrase. HIV-1 subtype was determined using the REGA HIV-1 subtyping tool. Resistance mutations were interpreted according to the Stanford HIV drug resistance database ( http://hivdb.stanford.edu ) and the updated International Antiviral Society (IAS)-USA mutation lists. Moreover, rates of polymorphisms in the current isolates were compared with South African and global HIV-1C isolates. RESULTS: All subjects were infected with HIV-1C concordant to the protease (PR) and reverse transcriptase (RT) regions. Neither major resistance-associated IN mutations (T66I/A/K, E92Q/G, T97A, Y143HCR, S147G, Q148H/R/K, and N155H) nor silent mutations known to change the genetic barrier were observed. Moreover, the DDE-catalytic motif (D64G/D116G/E152 K) and signature HHCC zinc-binding motifs at codon 12, 16, 40 and 43 were found to be highly conserved. However, compared to other South African subtype C isolates, the rate of polymorphism was variable at various positions. CONCLUSION: Although the sample size is small, the findings suggest that this drug class could be effective in Ethiopia and other southern African countries where HIV-1C is predominantly circulating. The data will contribute to define the importance of integrase polymorphism and to improve resistance interpretation algorithms in HIV-1C isolates.


Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Farmacorresistencia Viral/genética , Inhibidores de Integrasa VIH/farmacología , VIH-1/efectos de los fármacos , Mutación , VIH-1/genética , Humanos , ARN Viral/sangre
8.
BMC Infect Dis ; 14: 158, 2014 Mar 22.
Artículo en Inglés | MEDLINE | ID: mdl-24655349

RESUMEN

BACKGROUND: Despite the increasing use of antiretroviral treatment (ART) recent data on frequency and pattern of drug resistance mutations in Ethiopia is not available. Furthermore with increasing mobility of people HIV-1 subtypes other than the predominant subtype C may likely be introduced from the neighbouring countries. This study was aimed to determine the molecular characterization and pre-antiretroviral treatment resistance mutations among HIV-1 chronically infected ART naïve patients after the roll out of ART in Ethiopia. METHODS: Viral RNA was determined in 160 baseline plasma samples. The entire PR and the first 335 codons (76%) of the RT regions of the pol gene of the HIV-1 genome (N = 160) were amplified and sequenced using an in-house assay. Genotypic drug resistance was defined as the presence of one or more resistance-related mutations as specified by the consensus mutation of Stanford University HIVDB and the International Antiviral Society (IAS) mutation lists. RESULTS: A predominance of HIV-1 subtype C (98.7%) was observed. The level of drug resistance is found to be 5.6% and 13.1% according to the Stanford University HIVDB drug resistance interpretation algorithms and the International Antiviral Society mutation lists, respectively. Mutations conferring simultaneous resistance to NRTIs and NNRTIs were not detected and no major PR mutation was found. However, a high rate of polymorphic changes both in PR and RT regions were observed. Moreover, twenty four (15%) monophyletic transmission clusters with bootstrap value of 99% were found. CONCLUSIONS: Strong evidence for consistent HIV-1C clade homogeneity and low influx of other variant into the country was found. The level of drug resistance observed in chronically infected treatment naïve patients which exceeds the WHO estimates suggests the need for incorporation of HIV-1 drug resistance testing prior to ART initiation. The occurrence of monophyletic transmission clusters affecting (24/160) individuals indicates their potential risk related practice. Thus, an intensified public health intervention program and monitoring of HIV drug resistance testing appears indispensible.


Asunto(s)
Antirretrovirales/uso terapéutico , Farmacorresistencia Viral/genética , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/virología , VIH-1/genética , Productos del Gen pol del Virus de la Inmunodeficiencia Humana/genética , Adulto , Etiopía/epidemiología , Femenino , Infecciones por VIH/epidemiología , Humanos , Masculino , Datos de Secuencia Molecular , Mutación , Polimorfismo Genético
9.
BMC Infect Dis ; 14: 28, 2014 Jan 14.
Artículo en Inglés | MEDLINE | ID: mdl-24422906

RESUMEN

BACKGROUND: Introduction of antiretroviral therapy (ART) in sub-Saharan Africa was a hot debate due to many concerns about adherence, logistics and resistance. Currently, it has been significantly scaled up. However as the WHO clinico-immunological approaches for initiation and monitoring of ART in the region lacks viral load determination and drug resistance monitoring, HIV infected adults and children may be at risk for "unrecognized" virologic failure and the subsequent development of antiretroviral drug resistance. This study evaluates the virological efficacy and immunological recovery of HIV/AIDS patients under ART. METHODS: Consecutive HIV-1 infected adults (N = 100) and children (N = 100) who have been receiving ART for up to 6 years at Gondar University Hospital, Ethiopia were enrolled following the WHO protocol for assessment of acquired drug resistance. Magnitude of viral suppression, genotypic drug resistance mutations and patterns of CD4+ T cell recovery were determined using standard virological and immunological methods. RESULTS: Virological suppression (HIV RNA < 40 copies/ml) was observed in 82 and 87% of adults and children on a median time of 24 months on ART, respectively. Mutation K103N conferring resistance to non nucleoside reverse transcriptase inhibitors and thymidine analogue mutations (M41L, L210W) were found only in one adult and child patient, respectively. Median CD4+ T cell count has increased from baseline 124 to 266 (IQR: 203-306) and 345 (IQR: 17-1435) to 998 (IQR: 678-2205) cells/mm3 in adults and children respectively after 12 months of ART. Nevertheless, small but significant number of clinically asymptomatic adults (16%) and children (13%) had low level viraemia (HIV-1 RNA 41-1000 copies/ml). CONCLUSIONS: Majority of both adults (82%) and children (87%) who received ART showed high viral suppression and immunological recovery. This indicates that despite limited resources in the setting virological efficacy can be sustained for a substantial length of time and also enhance immunological recovery irrespective of age. However, the presence of drug resistance mutations and low level viraemia among clinically asymptomatic patients highlights the need for virological monitoring.


Asunto(s)
Antirretrovirales/uso terapéutico , Linfocitos T CD4-Positivos , Infecciones por VIH/tratamiento farmacológico , Carga Viral , Adulto , Recuento de Linfocito CD4 , Niño , Preescolar , Farmacorresistencia Viral/genética , Etiopía , Femenino , Genotipo , VIH-1/genética , VIH-1/aislamiento & purificación , Humanos , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Mutación , Inhibidores de la Transcriptasa Inversa/uso terapéutico , Viremia/tratamiento farmacológico
11.
Viruses ; 14(8)2022 08 20.
Artículo en Inglés | MEDLINE | ID: mdl-36016451

RESUMEN

Rhinoviruses (RVs) constitute a substantial public health burden. To evaluate their abundance and genetic diversity in pediatric patients, RV RNA in respiratory samples was assessed using real-time RT-PCR and partial nucleic acid sequencing of viral genomes. Additionally, clinical data were retrieved from patient charts to determine the clinical significance of pediatric RV infections. In total, the respiratory specimens of 776 patients (<18 years), collected from 2013 to 2017, were analyzed. Infections occurred throughout the entire year, with peaks occurring in fall and winter, and showed remarkably high intra- and interseasonal diversity for RV genotypes. RV species were detected in the following frequencies: 49.1% RV-A, 5.9% RV-B, and 43.6% RV-C. RV-C was found to be more frequently associated with asthma (p = 0.04) and bronchiolitis (p < 0.001), while RV-A was more frequently associated with fever (p = 0.001) and pneumonia (p = 0.002). Additionally, 35.3% of the patients had co-infections with other pathogens, which were associated with a longer hospital stay (p < 0.001), need for ventilation (p < 0.001), and pneumonia (p < 0.001). Taken together, this study shows pronounced RV genetic diversity in pediatric patients and indicates differences in RV-associated pathologies, as well as an important role for co-infections.


Asunto(s)
Coinfección , Enterovirus , Infecciones por Picornaviridae , Infecciones del Sistema Respiratorio , Niño , Enterovirus/genética , Humanos , Epidemiología Molecular , Infecciones del Sistema Respiratorio/epidemiología , Rhinovirus/genética , Centros de Atención Terciaria
12.
Viruses ; 14(6)2022 06 14.
Artículo en Inglés | MEDLINE | ID: mdl-35746772

RESUMEN

Despite available vaccines, antibodies and antiviral agents, the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) pandemic still continues to cause severe disease and death. Current treatment options are limited, and emerging new mutations are a challenge. Thus, novel treatments and measures for prevention of viral infections are urgently required. Photodynamic inactivation (PDI) is a potential treatment for infections by a broad variety of critical pathogens, including viruses. We explored the infectiousness of clinical SARS-CoV-2 isolates in Vero cell cultures after PDI-treatment, using the photosensitizer Tetrahydroporphyrin-tetratosylate (THPTS) and near-infrared light. Replication of viral RNA (qPCR), viral cytopathic effects (microscopy) and mitochondrial activity were assessed. PDI of virus suspension with 1 µM THPTS before infection resulted in a reduction of detectable viral RNA by 3 log levels at day 3 and 6 after infection to similar levels as in previously heat-inactivated virions (<99.9%; p < 0.05). Mitochondrial activity, which was significantly reduced by viral infection, was markedly increased by PDI to levels similar to uninfected cell cultures. When applying THPTS-based PDI after infection, a single treatment had a virus load-reducing effect only at a higher concentration (3 µM) and reduced cell viability in terms of PDI-induced toxicity. Repeated PDI with 0.3 µM THPTS every 4 h for 3 d after infection reduced the viral load by more than 99.9% (p < 0.05), while cell viability was maintained. Our data demonstrate that THPTS-based antiviral PDI might constitute a promising approach for inactivation of SARS-CoV-2. Further testing will demonstrate if THPTS is also suitable to reduce the viral load in vivo.


Asunto(s)
Tratamiento Farmacológico de COVID-19 , SARS-CoV-2 , Animales , Antivirales/farmacología , Chlorocebus aethiops , Pandemias , ARN Viral/genética , Células Vero
13.
J Med Virol ; 83(5): 776-82, 2011 May.
Artículo en Inglés | MEDLINE | ID: mdl-21351106

RESUMEN

The prevalence of different genotypes of hepatitis C virus (HCV) in Ethiopia is not known. HCV genotypes influence the response to therapy with alpha-interferon alone or in combination with ribavirin. A cross sectional study was conducted on attendees of voluntary counseling and testing center. Serum samples from 1,954 (734 HIV positive and 1,220 HIV negative) individuals were screened for HCV antibody. Active HCV infection was confirmed by quantitative PCR in 18 of the 71 samples with anti-HCV antibodies. The HCV viral load ranged from 39,650 to 9,878,341 IU/ml (median 1,589,631 IU/ml) with no significant difference [χ(2)(17) = 18.00, P = 0.389] between persons positive or negative for HIV. The viral load of HCV was, however, higher in older study subjects (r = 0.80, P = 0.000). HCV genotypes were determined using the VERSANT HCV Genotype Assay (LiPA) and sequence analysis of the NS5b region of the HCV genome. Diverse HCV genotypes were found including genotypes 1, 2, 4, and 5. There was no difference in the distribution regarding the HIV status. As in other parts of the world, genotyping of HCV must be considered whenever HCV is incriminated as a cause of hepatitis.


Asunto(s)
Hepacivirus/clasificación , Hepacivirus/aislamiento & purificación , Hepatitis C/epidemiología , Hepatitis C/virología , Carga Viral , Adulto , Anciano , Consejo , Estudios Transversales , Etiopía/epidemiología , Femenino , Genotipo , Hepacivirus/genética , Anticuerpos contra la Hepatitis C/sangre , Humanos , Masculino , Persona de Mediana Edad , Tipificación Molecular , Reacción en Cadena de la Polimerasa , Prevalencia , ARN Viral/genética , Análisis de Secuencia de ADN , Proteínas no Estructurales Virales/genética
14.
Anal Biochem ; 411(1): 164-6, 2011 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-21094121

RESUMEN

Guanidinium thiocyanate-phenol-chloroform extraction (GTPC extraction) is widely used in molecular biology for isolating DNA, RNA, and proteins. Protein isolation by commercially available GTPC solutions is time consuming and the resulting pellets are only incompletely soluble. In this study ethanol-bromochloropropane-water was used for precipitation of proteins from the phenol-ethanol phase after GTPC extraction of RNA and DNA. The precipitated proteins can be readily dissolved in 4% SDS for subsequent analysis. This technique allows a fast (30min) and efficient (with a protein recovery of up to 95%) extraction of proteins for the study of transcriptional and posttranscriptional events from the same sample.


Asunto(s)
Bioquímica/métodos , Ácidos Nucleicos/aislamiento & purificación , Proteínas/aislamiento & purificación , Animales , Chlorocebus aethiops , Electroforesis en Gel de Poliacrilamida , Células HEK293 , Humanos , Ratas , Células Vero
15.
Diagn Microbiol Infect Dis ; 99(3): 115244, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33253961

RESUMEN

Nosocomial virus infections cause significant morbidity and mortality. Besides influenza viruses, the disease burden of parainfluenza virus type 3 (PIV-3) is comparatively high among hospitalized patients and severe disease courses can occur. PIV-3 showed the highest rates of nosocomial infections of a panel of respiratory viruses. Therefore, a retrospective observational study was conducted among patients with either PIV-3 or influenza viruses, which served as reference pathogen. The aim was to compare the seasonal dynamics and clinical characteristics of nosocomial infections with these highly transmittable viruses. Nosocomial infection occurred in 15.8% (n = 177) of all influenza cases, mainly in the first half of a season. About 24.3% (n = 104) of the PIV-3 cases were nosocomial and occurred mainly in the second half of a season. Both nosocomial rates of influenza and nosocomial rates of PIV-3 varied between the seasons. Community acquired and nosocomial cases differed in underlying medical conditions and immunosuppression. Knowledge of the baseline rates of nosocomial infections could contribute to the implementation of appropriate infection control measures.


Asunto(s)
Infección Hospitalaria/epidemiología , Infección Hospitalaria/virología , Hospitales Universitarios/estadística & datos numéricos , Gripe Humana/epidemiología , Infecciones por Respirovirus/epidemiología , Adolescente , Adulto , Niño , Preescolar , Infección Hospitalaria/transmisión , Femenino , Alemania/epidemiología , Humanos , Lactante , Gripe Humana/transmisión , Masculino , Persona de Mediana Edad , Orthomyxoviridae/patogenicidad , Virus de la Parainfluenza 3 Humana/patogenicidad , Infecciones del Sistema Respiratorio/virología , Infecciones por Respirovirus/transmisión , Estudios Retrospectivos , Estaciones del Año , Adulto Joven
16.
Viruses ; 13(10)2021 10 08.
Artículo en Inglés | MEDLINE | ID: mdl-34696457

RESUMEN

Rhinoviruses (RVs) constitute a substantial public health burden. To evaluate their abundance and genetic diversity in adult patients, RV RNA in respiratory samples was assessed using real-time RT-PCR and the partial nucleic acid sequencing of viral genomes. Additionally, clinical data were retrieved from patient charts to determine the clinical significance of adult RV infections. In total, the respiratory specimens of 284 adult patients (18-90 years), collected from 2013 to 2017, were analyzed. Infections occurred throughout the entire year, with peaks occurring in fall and winter, and showed a remarkably high intra- and interseasonal diversity of RV genotypes. RV species were detected in the following ratios: 60.9% RV-A 173, 12.7% RV-B, and 26.4% RV-C. No correlations between RV species and underlying comorbidities such as asthma (p = 0.167), COPD (p = 0.312) or immunosuppression (p = 0.824) were found. However, 21.1% of the patients had co-infections with other pathogens, which were associated with a longer hospital stay (p = 0.024), LRTI (p < 0.001), and pneumonia (p = 0.01). Taken together, this study shows a pronounced genetic diversity of RV in adults and underlines the important role of co-infections. No correlation of specific RV species with a particular clinical presentation could be deduced.


Asunto(s)
Epidemiología Molecular , Infecciones por Picornaviridae/epidemiología , Infecciones por Picornaviridae/virología , Rhinovirus/genética , Centros de Atención Terciaria , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Coinfección/epidemiología , Coinfección/virología , Femenino , Genoma Viral , Genotipo , Alemania/epidemiología , Humanos , Masculino , Persona de Mediana Edad , Filogenia , Infecciones por Picornaviridae/diagnóstico , Salud Pública , Reacción en Cadena en Tiempo Real de la Polimerasa , Sistema Respiratorio/virología , Infecciones del Sistema Respiratorio/virología , Rhinovirus/clasificación , Rhinovirus/aislamiento & purificación , Estaciones del Año , Adulto Joven
17.
Viruses ; 13(9)2021 08 30.
Artículo en Inglés | MEDLINE | ID: mdl-34578303

RESUMEN

Reverse genetics is a technology that allows the production of a virus from its complementary DNA (cDNA). It is a powerful tool for analyzing viral genes, the development of novel vaccines, and gene delivery vectors. The standard reverse genetics protocols are laborious, time-consuming, and inefficient for negative-strand RNA viruses. A new reverse genetics platform was established, which increases the recovery efficiency of the measles virus (MV) in human 293-3-46 cells. The novel features compared with the standard system involving 293-3-46 cells comprise (a) dual promoters containing the RNA polymerase II promoter (CMV) and the bacteriophage T7 promoter placed in uni-direction on the same plasmid to enhance RNA transcription; (b) three G nucleotides added just after the T7 promoter to increase the T7 RNA polymerase activity; and (c) two ribozymes, the hairpin hammerhead ribozyme (HHRz), and the hepatitis delta virus ribozyme (HDVrz), were used to cleavage the exact termini of the antigenome RNA. Full-length antigenome cDNA of MV of the wild type IC323 strain or the vaccine AIK-C strain was inserted into the plasmid backbone. Both virus strains were easily rescued from their respective cloned cDNA. The rescue efficiency increased up to 80% compared with the use of the standard T7 rescue system. We assume that this system might be helpful in the rescue of other human mononegavirales.


Asunto(s)
Virus del Sarampión/genética , Regiones Promotoras Genéticas , Virus Reordenados/genética , Genética Inversa/métodos , Animales , Bacteriófago T7/genética , Chlorocebus aethiops , ADN Complementario , ARN Polimerasas Dirigidas por ADN/metabolismo , Genoma Viral , Humanos , ARN Viral/genética , Células Vero , Proteínas Virales/metabolismo
18.
Diagn Microbiol Infect Dis ; 100(4): 115382, 2021 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-33930691

RESUMEN

Sensitivity and specificity of serological assays are key parameters for the accurate estimation of SARS-CoV-2 sero-prevalence. The aim of this study was to compare 8 readily available IgG antibody tests using a panel of well-defined serum samples of prepandemic and pandemic origin. A cross-reaction panel included samples of patients with recent infection with either of the endemic Coronaviruses 229E, NL63, HKU1, or OC43. Additionally, samples with high antibody levels against influenza virus, adenovirus, and during acute EBV infection were included. Previous infection with endemic coronaviruses caused a significant amount of cross-reactivity in two of the assays. In contrast, the confidence intervals for the assays of Abbott, DiaSorin, Euroimmun and Roche encompassed the value of 98% for samples with a previous endemic HCoV infection. For all assays, sensitivities were between 91.3% and 98.8%. Assay performance was independent of the usage of either nucleocapsid or spike proteins.


Asunto(s)
Anticuerpos Antivirales/sangre , Prueba Serológica para COVID-19/métodos , COVID-19/diagnóstico , Inmunoglobulina G/sangre , SARS-CoV-2/inmunología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , COVID-19/inmunología , Prueba Serológica para COVID-19/normas , Niño , Femenino , Humanos , Masculino , Persona de Mediana Edad , Sensibilidad y Especificidad , Estudios Seroepidemiológicos , Proteínas Virales , Adulto Joven
19.
Nat Biotechnol ; 39(6): 705-716, 2021 06.
Artículo en Inglés | MEDLINE | ID: mdl-33361824

RESUMEN

In coronavirus disease 2019 (COVID-19), hypertension and cardiovascular diseases are major risk factors for critical disease progression. However, the underlying causes and the effects of the main anti-hypertensive therapies-angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin receptor blockers (ARBs)-remain unclear. Combining clinical data (n = 144) and single-cell sequencing data of airway samples (n = 48) with in vitro experiments, we observed a distinct inflammatory predisposition of immune cells in patients with hypertension that correlated with critical COVID-19 progression. ACEI treatment was associated with dampened COVID-19-related hyperinflammation and with increased cell intrinsic antiviral responses, whereas ARB treatment related to enhanced epithelial-immune cell interactions. Macrophages and neutrophils of patients with hypertension, in particular under ARB treatment, exhibited higher expression of the pro-inflammatory cytokines CCL3 and CCL4 and the chemokine receptor CCR1. Although the limited size of our cohort does not allow us to establish clinical efficacy, our data suggest that the clinical benefits of ACEI treatment in patients with COVID-19 who have hypertension warrant further investigation.


Asunto(s)
Tratamiento Farmacológico de COVID-19 , Quimiocina CCL3/genética , Quimiocina CCL4/genética , Hipertensión/tratamiento farmacológico , Receptores CCR1/genética , Adulto , Antagonistas de Receptores de Angiotensina/administración & dosificación , Antagonistas de Receptores de Angiotensina/efectos adversos , Inhibidores de la Enzima Convertidora de Angiotensina/administración & dosificación , Inhibidores de la Enzima Convertidora de Angiotensina/efectos adversos , COVID-19/complicaciones , COVID-19/genética , COVID-19/virología , Progresión de la Enfermedad , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Hipertensión/complicaciones , Hipertensión/genética , Hipertensión/patología , Inflamación/complicaciones , Inflamación/tratamiento farmacológico , Inflamación/genética , Inflamación/virología , Masculino , Persona de Mediana Edad , RNA-Seq , Sistema Respiratorio/efectos de los fármacos , Sistema Respiratorio/patología , Sistema Respiratorio/virología , Factores de Riesgo , SARS-CoV-2/patogenicidad , Análisis de la Célula Individual
20.
Clin Oral Investig ; 14(1): 11-7, 2010 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-20054591

RESUMEN

Since the spring of 2009, there have been a considerable number of infected as well as fatal cases by virologically confirmed swine-origin H1N1 influenza A virus (S-OIV). The virus continues to spread globally. The World Health Organization (WHO) has now raised the level of S-OIV influenza pandemic alert to phase 6 ('the pandemic phase') because of the human-to-human transmission of the virus and the community-level outbreaks worldwide. The WHO also issues its concerns about the global surveillance, the diagnostic capacity for the infection and the pandemic preparedness plan in every country. However, no critical review on S-OIV influenza and dental practice published in the literature exists hitherto. Based on information up to November 2009, the aim of this article was to summarise significant data on this novel virus and a clinical practice guideline for dental professionals.


Asunto(s)
Subtipo H1N1 del Virus de la Influenza A , Gripe Humana , Antivirales/uso terapéutico , Inhibidores Enzimáticos/uso terapéutico , Humanos , Control de Infección Dental/métodos , Subtipo H1N1 del Virus de la Influenza A/inmunología , Vacunas contra la Influenza , Gripe Humana/prevención & control , Gripe Humana/virología , Neuraminidasa/antagonistas & inhibidores , Guías de Práctica Clínica como Asunto , Estados Unidos
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