RESUMEN
Systemic lupus erythematosus (SLE) has been associated with increased risk of tuberculosis (TB). However, little is known about the extent and risk factors for TB among Asian patient with SLE. We aimed to assess the rate of TB in patients with SLE, and investigate the risk of SLE on TB development using hospital administrative database. This is an historical cohort study of hospital discharge database from 2004 to 2011 to identify cases with SLE and TB using International Statistical Classification of Diseases and Related Health Problems, 9th Revision, Australian Modification (ICD-9-AM) codes. Of 301568 hospitalized patients, 841 (0.3%) patients had SLE, 1843 (0.6%) patients had TB, including 17 SLE patients (2.0%). SLE patients had a significantly higher rate of TB (2.0 vs. 0.6%, p < 0.001) compared to that of patients without SLE. The differences in the higher rate after breaking down was in the pulmonary TB group (1.7 vs. 0.5%, p < 0.00) but not in extrapulmonary TB group (0.4 vs. 0.1%, p = 0.060). Logistic regression analyses showed that SLE was a significant and independent predictor of TB (odds ratio 4.6, 95% CI 2.8-7.5, p < 0.001) after adjustment for factors such as age group, gender, ethnicity, admission class, nutritional deficiency, organ transplantation, and Charlson comorbidity index. SLE patients were found to experience higher rates of tuberculosis in this group of Asian patient population. Patients with SLE should be considered as a high-risk group for TB, active screening for latent patients and treatment for positive TB patients is needed.
Asunto(s)
Lupus Eritematoso Sistémico/epidemiología , Centros de Atención Terciaria , Tuberculosis/epidemiología , Anciano , Distribución de Chi-Cuadrado , Comorbilidad , Bases de Datos Factuales , Femenino , Humanos , Modelos Logísticos , Lupus Eritematoso Sistémico/diagnóstico , Masculino , Persona de Mediana Edad , Oportunidad Relativa , Pronóstico , Medición de Riesgo , Factores de Riesgo , Singapur/epidemiología , Tuberculosis/diagnósticoRESUMEN
PURPOSE: To report late rectal and bladder toxicity outcomes of a CT-based image-guided brachytherapy (IGBT) technique for treatment of cervical cancer. METHODS AND MATERIALS: Between 2008 and 2014, 95 women with International Federation of Gynecology and Obstetrics stage IB to IVA cervical carcinoma treated with definitive concurrent cisplatin-based chemotherapy and external beam radiation therapy 50.4 Gy in 28 fractions followed by planned prescription dose of 7 Gy × 4 fractions of high-dose-rate IGBT was retrospectively reviewed. At each implantation, all patients had a urinary catheter in situ and received bowel enema before undergoing planning CT simulation. A high-risk clinical target volume (HRCTV) as per GEC-ESTRO guidelines and the entire cervix, rectum, and bladder was contoured on the simulation CT according to Radiation Therapy Oncology Group Gynaecology Contouring Atlas. Reported doses to HRCTV and organs at risk were recorded. Toxicities were recorded using National Cancer Institute Common Terminology Criteria for Adverse Events version 3. RESULTS: The median followup time was 29 months. The mean HRCTV equivalent dose in 2 Gy fractions (EQD2) of external beam radiation therapy combined with brachytherapy was 80 Gy (standard deviation [SD], 11), and the rectal doses to 2 cm3 (D2cc) EQD2 and bladder D2cc EQD2 were 74 Gy (SD, 6) and 79 Gy (SD, 15), respectively. Twenty-two patients (23%) had grade 2 proctitis and 10 patients (11%) had grade 3 proctitis. Four patients (4%) had grade 2 cystitis and two patients (2%) had grade 3 cystitis. No patients had ≥ grade 4 toxicity. CONCLUSIONS: Despite CT-based brachytherapy planning, reported organ at risk toxicity was still significant compared with reported MRI-based planning series. Coimplementation of interstitial IGBT using the European Study on MRI-guided Brachytherapy in Locally Advanced Cervical Cancer (EMBRACE) protocol or using intensity-modulated radiation therapy during the external beam phase treatment might help to limit these late toxicities.
Asunto(s)
Adenocarcinoma/terapia , Braquiterapia/métodos , Carcinoma de Células Escamosas/terapia , Cistitis/epidemiología , Proctitis/epidemiología , Traumatismos por Radiación/epidemiología , Neoplasias del Cuello Uterino/terapia , Adenocarcinoma/diagnóstico por imagen , Adenocarcinoma/patología , Adulto , Anciano , Antineoplásicos/uso terapéutico , Braquiterapia/efectos adversos , Carcinoma de Células Escamosas/diagnóstico por imagen , Carcinoma de Células Escamosas/patología , Quimioradioterapia/métodos , Cisplatino/uso terapéutico , Colon Sigmoide/diagnóstico por imagen , Cistitis/etiología , Estudios de Factibilidad , Femenino , Humanos , Persona de Mediana Edad , Órganos en Riesgo , Proctitis/etiología , Traumatismos por Radiación/etiología , Dosificación Radioterapéutica , Planificación de la Radioterapia Asistida por Computador/métodos , Radioterapia de Intensidad Modulada/métodos , Recto/diagnóstico por imagen , Estudios Retrospectivos , Tomografía Computarizada por Rayos X/métodos , Vejiga Urinaria/diagnóstico por imagen , Neoplasias del Cuello Uterino/diagnóstico por imagen , Neoplasias del Cuello Uterino/patología , Adulto JovenRESUMEN
Cancer patients with Down syndrome (DS) are at increased risk for anthracycline-related cardiotoxicity. Mitochondrial DNA (mtDNA) alterations in hearts with-DS may contribute to anthracycline-related cardiotoxicity. Cardiac mtDNA and the mtDNA(4977) deletion were quantitated in samples with- (n = 11) and without-DS (n = 31). Samples with-DS showed 30% lower mtDNA (DS(MT-ND1/18Sratio): 1.48 ± 0.72 versus non-DS(MT-ND1/18Sratio): 2.10 ± 1.59; p = 0.647) and 30% higher frequency of the mtDNA(4977) deletion (DS(% frequency mtDNA(4977)) deletion: 0.0086 ± 0.0166 versus non-DS(% frequency mtDNA(4977)) deletion: 0.0066 ± 0.0124, p = 0.514) than samples without-DS. The BACH1 and microRNA-155 (miR-155) genes are located in chromosome 21, and their products have demonstrated roles during oxidative stress. BACH1 and miR-155 expression did not differ in hearts with- and without-DS. An association between BACH1 and miR-155 expression was detected in hearts without-DS, suggesting alterations between BACH1-miR-155 interactions in the DS settings.
Asunto(s)
Antraciclinas/efectos adversos , Antibióticos Antineoplásicos/efectos adversos , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , ADN Mitocondrial/genética , Síndrome de Down/genética , Proteínas del Grupo de Complementación de la Anemia de Fanconi/genética , MicroARNs/genética , Adolescente , Adulto , Anciano , Antraciclinas/uso terapéutico , Antibióticos Antineoplásicos/uso terapéutico , Secuencia de Bases , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/biosíntesis , Cardiotoxicidad , Niño , Preescolar , Cromosomas Humanos Par 21 , Proteínas del Grupo de Complementación de la Anemia de Fanconi/biosíntesis , Femenino , Genes Mitocondriales , Cardiopatías/inducido químicamente , Cardiopatías/genética , Cardiopatías/patología , Humanos , Lactante , Leucemia Mieloide Aguda/tratamiento farmacológico , Masculino , MicroARNs/biosíntesis , Persona de Mediana Edad , Mitocondrias/efectos de los fármacos , Miocardio/metabolismo , Estrés Oxidativo/genética , Estrés Oxidativo/fisiología , Proyectos Piloto , ARN Mensajero/genética , Análisis de Secuencia de ADN , Eliminación de Secuencia/genética , Adulto JovenRESUMEN
Many pollinator populations are declining, with large economic and ecological implications. Parasites are known to be an important factor in the some of the population declines of honey bees and bumblebees, but little is known about the parasites afflicting most other pollinators, or the extent of interspecific transmission or vectoring of parasites. Here we carry out a preliminary screening of pollinators (honey bees, five species of bumblebee, three species of wasp, four species of hoverfly and three genera of other bees) in the UK for parasites. We used molecular methods to screen for six honey bee viruses, Ascosphaera fungi, Microsporidia, and Wolbachia intracellular bacteria. We aimed simply to detect the presence of the parasites, encompassing vectoring as well as actual infections. Many pollinators of all types were positive for Ascosphaera fungi, while Microsporidia were rarer, being most frequently found in bumblebees. We also detected that most pollinators were positive for Wolbachia, most probably indicating infection with this intracellular symbiont, and raising the possibility that it may be an important factor in influencing host sex ratios or fitness in a diversity of pollinators. Importantly, we found that about a third of bumblebees (Bombus pascuorum and Bombus terrestris) and a third of wasps (Vespula vulgaris), as well as all honey bees, were positive for deformed wing virus, but that this virus was not present in other pollinators. Deformed wing virus therefore does not appear to be a general parasite of pollinators, but does interact significantly with at least three species of bumblebee and wasp. Further work is needed to establish the identity of some of the parasites, their spatiotemporal variation, and whether they are infecting the various pollinator species or being vectored. However, these results provide a first insight into the diversity, and potential exchange, of parasites in pollinator communities.
Asunto(s)
Abejas/parasitología , Himenópteros/parasitología , Parásitos/patogenicidad , Polinización , Avispas/parasitología , Animales , Abejas/virología , ADN Viral/genética , Himenópteros/virología , Virus de Insectos/genética , Virus de Insectos/aislamiento & purificación , Reacción en Cadena de la Polimerasa , Avispas/virologíaRESUMEN
An aborted mid-gestational male Steller sea lion fetus with an attached placenta was recovered on the floor of an open floating capture trap located off Norris Rock near Denman Island, British Columbia. Viral culture of the placenta demonstrated cytopathic effect. Although no specific signal was obtained in microarray experiments using RNA obtained from viral culture, elution and sequence analysis revealed the presence of a reovirus. Complete genome pyrosequencing led to the identification of an orthoreovirus that we have tentatively named Steller sea lion reovirus (SSRV). Phylogenetic analysis revealed similarities between SSRV and orthoreoviruses of birds, bats and other mammals that suggests potential for interspecies transmission.
Asunto(s)
Feto Abortado/virología , Genoma Viral , Orthoreovirus/aislamiento & purificación , ARN Viral/genética , Leones Marinos/virología , Animales , Colombia Británica , Análisis por Conglomerados , Femenino , Masculino , Datos de Secuencia Molecular , Filogenia , Placenta/virología , Embarazo , Análisis de Secuencia de ADNRESUMEN
Leanyer virus (LEAV), currently classified as a member of the genus Orthobunyavirus, in the family Bunyaviridae, was originally isolated from a pool of Anopheles meraukensis mosquitoes, collected at Leanyer, Northern Territory, Australia in 1974. When it failed to react in serological tests with antisera from other known viruses, full-length genomic sequencing was pursued to determine the relationship of LEAV to other orthobunyavirus species. Genetic and serological characterization confirmed its antigenic distance from other orthobunyaviruses, including to its closest genetic neighbours, the Simbu group viruses, suggesting that it may represent a new antigenic complex.
Asunto(s)
Anopheles/virología , Orthobunyavirus/clasificación , Orthobunyavirus/genética , Filogenia , Animales , Australia , Genoma Viral , Genómica , Datos de Secuencia Molecular , Orthobunyavirus/aislamiento & purificaciónRESUMEN
While worldwide pandemic influenza A(H1N1) pdm case fatality rate (CFR) was 0.4%, Argentina's was 4.5%. A total of 34 strains from mild and severe cases were analyzed. A full genome sequencing was carried out on 26 of these, and a partial sequencing on the remaining eight. We observed no evidence that the high CFR can be attributed to direct virus changes. No evidence of re-assortment, mutations associated with resistance to antiviral drugs, or genetic drift that might contribute to virulence was observed. Although the mutation D225G associated with severity in the latest reports from the Ukraine and Norway is not observed among the Argentine strains, an amino acid change in the area (S206T) surrounding the HA receptor binding domain was observed, the same previously established worldwide.
Asunto(s)
ADN Viral/genética , Subtipo H1N1 del Virus de la Influenza A/genética , Gripe Humana/virología , Mutación/genética , Adolescente , Adulto , Argentina/epidemiología , Niño , Preescolar , Análisis por Conglomerados , Femenino , Humanos , Lactante , Subtipo H1N1 del Virus de la Influenza A/aislamiento & purificación , Gripe Humana/mortalidad , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , ARN Viral/genética , Receptores Virales/genética , Reproducibilidad de los Resultados , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
While worldwide pandemic influenza A(H1N1) pdm case fatality rate (CFR) was 0.4%, Argentina's was 4.5%. A total of 34 strains from mild and severe cases were analyzed. A full genome sequencing was carried out on 26 of these, and a partial sequencing on the remaining eight. We observed no evidence that the high CFR can be attributed to direct virus changes. No evidence of re-assortment, mutations associated with resistance to antiviral drugs, or genetic drift that might contribute to virulence was observed. Although the mutation D225G associated with severity in the latest reports from the Ukraine and Norway is not observed among the Argentine strains, an amino acid change in the area (S206T) surrounding the HA receptor binding domain was observed, the same previously established worldwide.
Mientras que la tasa de letalidad (CFR) para (H1N1)pdm en todo el mundo era del 0.4%, en la Argentina la mortalidad observada fue de 4.5%. La secuenciación del genoma completo de 26 cepas de virus argentinos de influenza A (H1N1)pdm de casos leves y graves y de 8 cepas secuenciadas parcialmente no mostró evidencia de que la elevada tasa de letalidad se pueda atribuir directamente a cambios en el virus. No se encontraron hallazgos de recombinación, de mutaciones asociadas con la resistencia a los medicamentos antivirales ni de variaciones genéticas que puedan contribuir a la virulencia observada. Si bien la mutación D225G asociada con la gravedad, comunicada en informes procedentes de Ucrania y Noruega, no se ha encontrado en las cepas argentinas estudiadas, se ha observado un cambio aminoacídico en la región (S206T) en torno al dominio del sitio de unión al receptor en la HA, el mismo hallado en cepas distribuidas alrededor del mundo.
Asunto(s)
Adolescente , Adulto , Niño , Preescolar , Femenino , Humanos , Lactante , Masculino , Persona de Mediana Edad , Adulto Joven , ADN Viral/genética , Subtipo H1N1 del Virus de la Influenza A/genética , Gripe Humana/virología , Mutación/genética , Argentina/epidemiología , Análisis por Conglomerados , Subtipo H1N1 del Virus de la Influenza A/aislamiento & purificación , Gripe Humana/mortalidad , Datos de Secuencia Molecular , Reproducibilidad de los Resultados , ARN Viral/genética , Receptores Virales/genética , Índice de Severidad de la EnfermedadRESUMEN
Atlantic salmon (Salmo salar L.) mariculture has been associated with epidemics of infectious diseases that threaten not only local production, but also wild fish coming into close proximity to marine pens and fish escaping from them. Heart and skeletal muscle inflammation (HSMI) is a frequently fatal disease of farmed Atlantic salmon. First recognized in one farm in Norway in 1999, HSMI was subsequently implicated in outbreaks in other farms in Norway and the United Kingdom. Although pathology and disease transmission studies indicated an infectious basis, efforts to identify an agent were unsuccessful. Here we provide evidence that HSMI is associated with infection with piscine reovirus (PRV). PRV is a novel reovirus identified by unbiased high throughput DNA sequencing and a bioinformatics program focused on nucleotide frequency as well as sequence alignment and motif analyses. Formal implication of PRV in HSMI will require isolation in cell culture and fulfillment of Koch's postulates, or prevention or modification of disease through use of specific drugs or vaccines. Nonetheless, as our data indicate that a causal relationship is plausible, measures must be taken to control PRV not only because it threatens domestic salmon production but also due to the potential for transmission to wild salmon populations.
Asunto(s)
Enfermedades de los Peces/virología , Cardiopatías/virología , Inflamación/inmunología , Inflamación/virología , Músculo Esquelético/virología , Reoviridae/patogenicidad , Salmo salar/virología , Animales , Femenino , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/patología , Cardiopatías/inmunología , Cardiopatías/patología , Músculo Esquelético/inmunología , Músculo Esquelético/patología , Reoviridae/inmunologíaRESUMEN
Lymphocytic choriomeningitis virus (LCMV) is the prototype of the family Arenaviridae. LCMV can be associated with severe disease in humans, and its global distribution reflects the broad dispersion of the primary rodent reservoir, the house mouse (Mus musculus). Recent interest in the natural history of the virus has been stimulated by increasing recognition of LCMV infections during pregnancy, and in clusters of LCMV-associated fatal illness among tissue transplant recipients. Despite its public health importance, little is known regarding the genetic diversity or distribution of virus variants. Genomic analysis of 29 LCMV strains collected from a variety of geographic and temporal sources showed these viruses to be highly diverse. Several distinct lineages exist, but there is little correlation with time or place of isolation. Bayesian analysis estimates the most recent common ancestor to be 1,000-5,000 years old, and this long history is consistent with complex phylogeographic relationships of the extant virus isolates.
Asunto(s)
Virus de la Coriomeningitis Linfocítica/genética , Animales , Teorema de Bayes , Femenino , Variación Genética , Humanos , Virus de la Coriomeningitis Linfocítica/clasificación , Ratones/virología , Persona de Mediana EdadRESUMEN
BACKGROUND: The analysis of oligonucleotide microarray data in pathogen surveillance and discovery is a challenging task. Target template concentration, nucleic acid integrity, and host nucleic acid composition can each have a profound effect on signal distribution. Exploratory analysis of fluorescent signal distribution in clinical samples has revealed deviations from normality, suggesting that distribution-free approaches should be applied. RESULTS: Positive predictive value and false positive rates were examined to assess the utility of three well-established nonparametric methods for the analysis of viral array hybridization data: (1) Mann-Whitney U, (2) the Spearman correlation coefficient and (3) the chi-square test. Of the three tests, the chi-square proved most useful. CONCLUSIONS: The acceptance of microarray use for routine clinical diagnostics will require that the technology be accompanied by simple yet reliable analytic methods. We report that our implementation of the chi-square test yielded a combination of low false positive rates and a high degree of predictive accuracy.
Asunto(s)
Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Virus/genética , Animales , Perfilación de la Expresión Génica , Humanos , Modelos Estadísticos , Hibridación de Ácido Nucleico/métodos , Estándares de ReferenciaRESUMEN
Merino Walk virus (MWV), a proposed novel tentative species of the family Arenaviridae, was isolated from a rodent, Myotomys unisulcatus, collected at Merino Walk, Eastern Cape, South Africa, in 1985. Full-length genomic sequence confirmed MWV as an arenavirus related distantly to Mobala, Mopeia and Ippy viruses, all members of the Old World arenavirus complex. We propose MWV as a tentative novel species in the Lassa-lymphocytic choriomeningitis virus complex, based on its isolation from a novel rodent species and its genetic and serological characteristics.
Asunto(s)
Infecciones por Arenaviridae/veterinaria , Arenavirus del Viejo Mundo/clasificación , Arenavirus del Viejo Mundo/patogenicidad , Genoma Viral , Murinae/virología , ARN Viral/genética , Animales , Animales Recién Nacidos , Infecciones por Arenaviridae/virología , Arenavirus del Viejo Mundo/aislamiento & purificación , Secuencia de Bases , Chlorocebus aethiops , Análisis por Conglomerados , Ratones , Ratones Endogámicos ICR , Datos de Secuencia Molecular , Conformación de Ácido Nucleico , Filogenia , Análisis de Secuencia de ADN , Homología de Secuencia , Sudáfrica , Células VeroRESUMEN
An enterovirus was cultured from an erosive tongue lesion of a bottlenose dolphin (Tursiops truncatus). The morphology of virions on negative staining electron microscopy was consistent with those of enteroviruses. Analysis of 2613 bp of the polyprotein gene identified the isolate as a novel enterovirus strain, tentatively named bottlenose dolphin enterovirus (BDEV), that nests within the species Bovine enterovirus. Serologic evidence of exposure to enteroviruses was common in both free-ranging and managed collection dolphins. Managed collection dolphins were more likely to have high antibody levels, although the highest levels were reported in free-ranging populations. Associations between enterovirus antibody levels, and age, sex, complete blood counts, and clinical serum biochemistries were explored. Dolphins with higher antibody levels were more likely to be hyperproteinemic and hyperglobulinemic.
Asunto(s)
Delfín Mular/virología , Infecciones por Enterovirus/veterinaria , Animales , Enterovirus Bovino/genética , Enterovirus Bovino/aislamiento & purificación , Ensayo de Inmunoadsorción Enzimática , Femenino , Datos de Secuencia Molecular , Filogenia , Alineación de Secuencia , Análisis de Secuencia de ADN , Lengua/virologíaRESUMEN
BACKGROUND: Initial reports in May 2009 of the novel influenza strain H1N1pdm estimated a case fatality rate (CFR) of 0.6%, similar to that of seasonal influenza. In July 2009, however, Argentina reported 3056 cases with 137 deaths, representing a CFR of 4.5%. Potential explanations for increased CFR included virus reassortment or genetic drift, or infection of a more vulnerable population. Virus genomic sequencing of 26 Argentinian samples representing both severe and mild disease indicated no evidence of reassortment, mutations associated with resistance to antiviral drugs, or genetic drift that might contribute to virulence. Furthermore, no evidence was found for increased frequency of risk factors for H1N1pdm disease. METHODS/PRINCIPAL FINDINGS: We examined nasopharyngeal swab samples (NPS) from 199 cases of H1N1pdm infection from Argentina with MassTag PCR, testing for 33 additional microbial agents. The study population consisted of 199 H1N1pdm-infected subjects sampled between 23 June and 4 July 2009. Thirty-nine had severe disease defined as death (n = 20) or hospitalization (n = 19); 160 had mild disease. At least one additional agent of potential pathogenic importance was identified in 152 samples (76%), including Streptococcus pneumoniae (n = 62); Haemophilus influenzae (n = 104); human respiratory syncytial virus A (n = 11) and B (n = 1); human rhinovirus A (n = 1) and B (n = 4); human coronaviruses 229E (n = 1) and OC43 (n = 2); Klebsiella pneumoniae (n = 2); Acinetobacter baumannii (n = 2); Serratia marcescens (n = 1); and Staphylococcus aureus (n = 35) and methicillin-resistant S. aureus (MRSA, n = 6). The presence of S. pneumoniae was strongly correlated with severe disease. S. pneumoniae was present in 56.4% of severe cases versus 25% of mild cases; more than one-third of H1N1pdm NPS with S. pneumoniae were from subjects with severe disease (22 of 62 S. pneumoniae-positive NPS, p = 0.0004). In subjects 6 to 55 years of age, the adjusted odds ratio (OR) of severe disease in the presence of S. pneumoniae was 125.5 (95% confidence interval [CI], 16.95, 928.72; p<0.0001). CONCLUSIONS/SIGNIFICANCE: The association of S. pneumoniae with morbidity and mortality is established in the current and previous influenza pandemics. However, this study is the first to demonstrate the prognostic significance of non-invasive antemortem diagnosis of S. pneumoniae infection and may provide insights into clinical management.
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Brotes de Enfermedades , Subtipo H1N1 del Virus de la Influenza A/fisiología , Gripe Humana/microbiología , Gripe Humana/patología , Infecciones Neumocócicas/complicaciones , Infecciones Neumocócicas/microbiología , Streptococcus pneumoniae/fisiología , Adolescente , Adulto , Argentina/epidemiología , Secreciones Corporales/microbiología , Secreciones Corporales/virología , Niño , Femenino , Humanos , Gripe Humana/complicaciones , Gripe Humana/epidemiología , Masculino , Persona de Mediana Edad , Infecciones Neumocócicas/virología , Reacción en Cadena de la Polimerasa , Factores de Riesgo , Adulto JovenRESUMEN
BACKGROUND: Respiratory infections are the most common infectious diseases in humans worldwide and are a leading cause of death in children less than 5 years of age. OBJECTIVES: Identify candidate pathogens in pediatric patients with unexplained respiratory disease. STUDY DESIGN: Forty-four nasopharyngeal washes collected during the 2004-2005 winter season from pediatric patients with respiratory illnesses that tested negative for 7 common respiratory pathogens by culture and direct immunofluorescence assays were analyzed by MassTag-PCR. To distinguish human enteroviruses (HEV) and rhinoviruses (HRV), samples positive for picornaviruses were further characterized by sequence analysis. RESULTS: Candidate pathogens were detected by MassTag PCR in 27 of the 44 (61%) specimens that previously were rated negative. Sixteen of these 27 specimens (59%) contained picornaviruses; of these 9 (57%) contained RNA of a recently discovered clade of rhinoviruses. Bocaviruses were detected in three patients by RT-PCR. CONCLUSIONS: Our study confirms that multiplex MassTag-PCR enhances the detection of pathogens in clinical specimens, and shows that previously unrecognized rhinoviruses, that potentially form a species HRV-C, may cause a significant amount of pediatric respiratory disease.
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Nasofaringe/virología , Infecciones por Picornaviridae/virología , Infecciones del Sistema Respiratorio , Rhinovirus , Virosis , Enfermedad Aguda , Preescolar , Enterovirus/aislamiento & purificación , Infecciones por Enterovirus/epidemiología , Infecciones por Enterovirus/virología , Femenino , Humanos , Lactante , Masculino , Infecciones por Picornaviridae/epidemiología , Reacción en Cadena de la Polimerasa/métodos , Prevalencia , Infecciones del Sistema Respiratorio/epidemiología , Infecciones del Sistema Respiratorio/virología , Rhinovirus/clasificación , Rhinovirus/aislamiento & purificación , Virosis/epidemiología , Virosis/virologíaRESUMEN
BACKGROUND: Three patients who received visceral-organ transplants from a single donor on the same day died of a febrile illness 4 to 6 weeks after transplantation. Culture, polymerase-chain-reaction (PCR) and serologic assays, and oligonucleotide microarray analysis for a wide range of infectious agents were not informative. METHODS: We evaluated RNA obtained from the liver and kidney transplant recipients. Unbiased high-throughput sequencing was used to identify microbial sequences not found by means of other methods. The specificity of sequences for a new candidate pathogen was confirmed by means of culture and by means of PCR, immunohistochemical, and serologic analyses. RESULTS: High-throughput sequencing yielded 103,632 sequences, of which 14 represented an Old World arenavirus. Additional sequence analysis showed that this new arenavirus was related to lymphocytic choriomeningitis viruses. Specific PCR assays based on a unique sequence confirmed the presence of the virus in the kidneys, liver, blood, and cerebrospinal fluid of the recipients. Immunohistochemical analysis revealed arenavirus antigen in the liver and kidney transplants in the recipients. IgM and IgG antiviral antibodies were detected in the serum of the donor. Seroconversion was evident in serum specimens obtained from one recipient at two time points. CONCLUSIONS: Unbiased high-throughput sequencing is a powerful tool for the discovery of pathogens. The use of this method during an outbreak of disease facilitated the identification of a new arenavirus transmitted through solid-organ transplantation.
Asunto(s)
Infecciones por Arenaviridae/virología , Arenavirus/clasificación , Trasplante de Riñón/efectos adversos , Trasplante de Hígado/efectos adversos , Análisis de Secuencia de ADN/métodos , Adulto , Anticuerpos Antivirales/sangre , Infecciones por Arenaviridae/transmisión , Arenavirus/genética , Arenavirus/aislamiento & purificación , Biología Computacional , Transmisión de Enfermedad Infecciosa , Femenino , Humanos , Inmunohistoquímica , Riñón/ultraestructura , Riñón/virología , Persona de Mediana Edad , Análisis de Secuencia por Matrices de Oligonucleótidos , Reacción en Cadena de la Polimerasa/métodos , ARN Viral/análisisRESUMEN
In colony collapse disorder (CCD), honey bee colonies inexplicably lose their workers. CCD has resulted in a loss of 50 to 90% of colonies in beekeeping operations across the United States. The observation that irradiated combs from affected colonies can be repopulated with naive bees suggests that infection may contribute to CCD. We used an unbiased metagenomic approach to survey microflora in CCD hives, normal hives, and imported royal jelly. Candidate pathogens were screened for significance of association with CCD by the examination of samples collected from several sites over a period of 3 years. One organism, Israeli acute paralysis virus of bees, was strongly correlated with CCD.
Asunto(s)
Bacterias/aislamiento & purificación , Abejas/microbiología , Abejas/virología , Genómica , Virus de Insectos/aislamiento & purificación , Nosema/aislamiento & purificación , Animales , Bacterias/clasificación , Bacterias/genética , Abejas/parasitología , Ácidos Grasos , Genes de ARNr , Virus de Insectos/clasificación , Virus de Insectos/genética , Nosema/clasificación , Nosema/genética , Filogenia , Virus ARN/clasificación , Virus ARN/genética , Virus ARN/aislamiento & purificación , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , ARN Viral/análisis , ARN Viral/genética , Análisis de Secuencia de ADN , Trypanosomatina/clasificación , Trypanosomatina/genética , Trypanosomatina/aislamiento & purificaciónRESUMEN
Acute respiratory infections are significant causes of morbidity, mortality, and economic burden worldwide. An accurate, early differential diagnosis may alter individual clinical management as well as facilitate the recognition of outbreaks that have implications for public health. Here we report on the establishment and validation of a comprehensive and sensitive microarray system for detection of respiratory viruses and subtyping of influenza viruses in clinical materials. Implementation of a set of influenza virus enrichment primers facilitated subtyping of influenza A viruses through the differential recognition of hemagglutinins 1 through 16 and neuraminidases 1 through 9. Twenty-one different respiratory virus species were accurately characterized, including a recently identified novel genetic clade of rhinovirus.
Asunto(s)
Virus de la Influenza A/clasificación , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Infecciones del Sistema Respiratorio/virología , Virus/aislamiento & purificación , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Humanos , Virus de la Influenza A/genética , Neuraminidasa/genética , Rhinovirus/clasificación , Rhinovirus/genética , Proteínas Virales/genética , Virus/genéticaRESUMEN
To facilitate rapid, unbiased, differential diagnosis of infectious diseases, we designed GreeneChipPm, a panmicrobial microarray comprising 29,455 sixty-mer oligonucleotide probes for vertebrate viruses, bacteria, fungi, and parasites. Methods for nucleic acid preparation, random primed PCR amplification, and labeling were optimized to allow the sensitivity required for application with nucleic acid extracted from clinical materials and cultured isolates. Analysis of nasopharyngeal aspirates, blood, urine, and tissue from persons with various infectious diseases confirmed the presence of viruses and bacteria identified by other methods, and implicated Plasmodium falciparum in an unexplained fatal case of hemorrhagic feverlike disease during the Marburg hemorrhagic fever outbreak in Angola in 2004-2005.