RESUMEN
We sought to clarify clinical features of exanthem subitum associated-encephalitis/encephalopathy, generally caused by primary human herpesvirus-6 infection in Japan. A two-part questionnaire was sent to hospitals between January 2003-December 2004. Of 3357 questionnaires, 2357 (70.2%) were returned, and 2293 (68.3%) were eligible for analysis. Eighty-six cases of exanthem subitum-associated encephalitis/encephalopathy were reported. Seventy-seven (89.5%) of 86 patients were diagnosed with human herpesvirus-6 infection by virologic examination. Although 41 (50.6%) of 81 patients had no sequelae, 38 (46.9%) had neurologic sequelae. Moreover, two fatal cases (2.5%) were reported. Pleocytosis was evident in only 4 (7.5%) of 53 patients, and cerebrospinal fluid protein levels were within normal range (23.4 +/- 14.6 mg/dL S.D.) in all patients. Human herpesvirus-6 DNA was detected in 21 (53.8%) of 39 patients. Abnormal computed tomography findings were a predictor of neurologic sequelae (P = 0.0097). As a consequence of this survey, we estimate that 61.9 cases of exanthem subitum-associated encephalitis occur every year. The disease prognosis was unexpectedly poor.
Asunto(s)
Encefalitis Viral/epidemiología , Exantema Súbito/epidemiología , Preescolar , ADN Viral , Encefalitis Viral/líquido cefalorraquídeo , Encefalitis Viral/complicaciones , Encefalitis Viral/patología , Exantema Súbito/líquido cefalorraquídeo , Exantema Súbito/complicaciones , Exantema Súbito/patología , Femenino , Herpesvirus Humano 6/genética , Herpesvirus Humano 6/aislamiento & purificación , Humanos , Lactante , Japón/epidemiología , Leucocitosis/líquido cefalorraquídeo , Leucocitosis/epidemiología , Leucocitosis/patología , Imagen por Resonancia Magnética , Masculino , Pronóstico , Encuestas y Cuestionarios , Tomografía Computarizada por Rayos XRESUMEN
OBJECTIVE: This study was conducted to examine the association between rotavirus antigenemia and clinical features, particularly extraintestinal manifestations, and the association between serum cytokine levels and rotavirus antigen quantity. METHODS: Sixty hospitalized children who received a diagnosis of acute rotavirus gastroenteritis were enrolled in this study. Paired serum samples were collected from the 60 children when admitted to and discharged from the hospital. Associations among viral antigen levels and fever, elevated transaminase levels, and seizures were evaluated to determine whether antigenemia correlated with disease severity. Viral antigen was measured by using an in-house enzyme-linked immunosorbent assay that detected VP6 antigen. A flow-cytometric bead array was used to measure serum cytokine levels. RESULTS: Rotavirus antigen levels were significantly higher in serum collected at the time of hospital admission than at the time of discharge. Serum rotavirus antigen levels peaked on day 2 of the illness (2.02 +/- 0.73), followed by a gradual decrease in antigen levels to nearly undetectable levels by day 6. The quantity of rotavirus antigen was significantly higher in serum collected from patients with fever than those without fever. The presence or absence of elevated transaminase levels and seizures was not associated with serum rotavirus antigen levels. A weak but significantly positive association was observed between interleukin 8 levels and antigenemia. A weak but significantly negative association was observed between interleukin 10 levels and antigenemia. CONCLUSIONS: Rotavirus antigenemia is frequently observed in a patient's serum during the acute phase, and viral antigen levels change dramatically during the acute phase of the illness. Because patients with fever had higher rotavirus antigen levels, antigenemia severity might contribute to fever. The host immune response plays an important role in controlling antigenemia levels.
Asunto(s)
Antígenos Virales/sangre , Citocinas/metabolismo , Gastroenteritis/inmunología , Gastroenteritis/virología , Infecciones por Rotavirus/inmunología , Rotavirus/inmunología , Enfermedad Aguda , Anticuerpos Antivirales/sangre , Biomarcadores/análisis , Niño , Preescolar , Estudios de Cohortes , Citocinas/análisis , Ensayo de Inmunoadsorción Enzimática , Heces/virología , Femenino , Gastroenteritis/terapia , Hospitalización/estadística & datos numéricos , Humanos , Japón , Masculino , Probabilidad , Medición de Riesgo , Rotavirus/aislamiento & purificación , Infecciones por Rotavirus/diagnóstico , Infecciones por Rotavirus/terapia , Sensibilidad y Especificidad , Índice de Severidad de la Enfermedad , Estadísticas no ParamétricasRESUMEN
BACKGROUND: A more rapid and easier method is needed for monitoring human herpesvirus 6 (HHV-6) infections. The loop-mediated isothermal amplification method (LAMP) can detect viral DNA with high specificity, efficiency, and speed under isothermal conditions. LAMP requires only simple equipment that is available in hospital laboratories. OBJECTIVES: We evaluated LAMP as a means of detecting HHV-6 DNA directly from patients' sera. RESULTS: The sensitivity of the HHV-6 LAMP protocol without heat denaturation was 1000 copies/tube; with heat denaturation 10 copies/tube were detected. Three hundred serum samples from children with fever were analyzed. Using HHV-6 isolation as a definition of HHV-6 infection, the sensitivity, specificity, positive predictive value, and negative predictive value of the HHV-6 LAMP method without DNA extraction were 95.5%, 95.2%, 94.0%, and 96.4%, respectively. CONCLUSION: Direct detection of HHV-6 DNA in serum with a modified HHV-6 LAMP could be used for rapid diagnosis of exanthem subitum (ES).
Asunto(s)
ADN Viral/sangre , Exantema Súbito/virología , Herpesvirus Humano 6/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Cartilla de ADN , Exantema Súbito/sangre , Femenino , Fiebre/sangre , Fiebre/virología , Humanos , Lactante , Masculino , Sensibilidad y EspecificidadRESUMEN
We present a case of pneumonia with marked pleural effusion caused by Aspergillus infection in a 2-year-old Japanese girl with Down's syndrome. The patient was previously diagnosed with acute myeloid leukemia, developing the pneumonia during induction treatment. Although no pathogens could be isolated from any clinical specimens, 135,000 copies/mL of Aspergillus DNA were detected in the pleural fluid using real time polymerase chain reaction. The copy numbers of DNA decreased rapidly after appropriate antifungal treatment.
Asunto(s)
Aspergilosis/complicaciones , Enfermedades Pulmonares Fúngicas/complicaciones , Derrame Pleural/etiología , Neumonía/complicaciones , Aspergilosis/microbiología , Aspergillus/genética , Aspergillus/aislamiento & purificación , Preescolar , ADN de Hongos/análisis , Femenino , Humanos , Enfermedades Pulmonares Fúngicas/microbiología , Derrame Pleural/microbiología , Neumonía/microbiología , Reacción en Cadena de la Polimerasa/métodosRESUMEN
To determine the cell populations in peripheral blood that are infected latently with human herpesvirus 7 (HHV-7), the real-time polymerase chain reaction (PCR) was used to determine the quantities of viral DNA in adherent and non-adherent cells from 71 healthy volunteers. Real-time PCR, which detected the U31 gene of HHV-7, was developed to measure viral load. The majority of non-adherent cells (14/16; 87.5%) contained HHV-7 DNA, while most of the adherent cells did not (1/16; 6.3%). HHV-7 viral load in non-adherent cells was significantly higher than that in adherent cells (P < 0.0001). Then, HHV-7 DNA load was compared between the CD4-positive and -negative cell fractions derived from the non-adherent cells of 26 healthy adults. As in the previous experiment, only 2 (7.7%) of the 26 adherent cell specimens contained small amounts of HHV-7 DNA (27.7 copies/1 x 10(6) cells and 208.7 copies/1 x 10(6) cells). In contrast, 88.5% of CD4(+) T cell samples (23/26 specimens) were positive for HHV-7 DNA, ranging from 0.4 to 3,542.8 copies/1 x 10(6) cells. Viral DNA was detected in only 3 (11.5%) of the 26 CD4(-) T cell specimens, with 8.4, 63.5, and 74.1 copies/1 x 10(6) cells. HHV-7-positive DNA loads were significantly higher in the CD4(+) T cells than those observed in the CD4(-) T cells (P = 0.0005). The relationship between HHV-7 viral loads in non-adherent cells and those in saliva was investigated. Comparison of HHV-7 DNA load between blood CD4(+) T cells and saliva revealed that the HHV-7 DNA load in saliva correlated with that present in CD4(+) T cells (r = 0.415; P = 0.0174).
Asunto(s)
Linfocitos T CD4-Positivos/virología , Herpesvirus Humano 7/aislamiento & purificación , Herpesvirus Humano 7/fisiología , Infecciones por Roseolovirus/virología , Latencia del Virus , Adulto , Adhesión Celular , ADN Viral/análisis , Femenino , Herpesvirus Humano 7/genética , Humanos , Masculino , Reacción en Cadena de la Polimerasa , Saliva/virología , Carga ViralRESUMEN
Primers for herpes simplex virus type 1 (HSV 1)-specific loop-mediated isothermal amplification (LAMP) method amplified HSV-1 DNA, while HSV-2-specific primers amplified only HSV-2 DNA; no LAMP products were produced by reactions performed with other viral DNAs. The sensitivities of the HSV-1- and HSV-2-specific LAMP methods, determined by agarose gel electrophoresis, reached 500 and 1,000 copies/tube, respectively. The turbidity assay, however, determined the sensitivity of the HSV-1- and HSV-2-specific LAMP methods to be 1,000 and 10,000 copies/tube, respectively. After initial validation studies, 18 swab samples (in sterilized water) collected from patients with either gingivostomatitis or vesicular skin eruptions were examined. HSV-1 LAMP products were detected by agarose gel electrophoresis in the 10 samples that also demonstrated viral DNA detection by real-time PCR. Nine of these 10 samples exhibited HSV-1 LAMP products by turbidity assay. Furthermore, both the agarose gel electrophoresis and the turbidity assay directly detected HSV-1 LAMP products in 9 of the 10 swab samples collected in sterilized water. Next, we examined the reliability of HSV type-specific LAMP for the detection of viral DNA in clinical specimens (culture medium) collected from genital lesions. HSV-2 was isolated from all of the samples and visualized by either agarose gel electrophoresis or turbidity assay.
Asunto(s)
Herpes Genital/diagnóstico , Herpes Simple/diagnóstico , Herpesvirus Humano 1/aislamiento & purificación , Herpesvirus Humano 2/aislamiento & purificación , Técnicas de Amplificación de Ácido Nucleico/métodos , ADN Viral/análisis , Herpes Genital/virología , Herpes Simple/virología , Herpesvirus Humano 1/genética , Herpesvirus Humano 2/genética , Humanos , Sensibilidad y Especificidad , Estomatitis Herpética/diagnóstico , Estomatitis Herpética/virología , Factores de TiempoRESUMEN
A case of neonatal human herpesvirus 6 (HHV-6) B infection is presented. Although HHV-6 B was isolated from peripheral blood at the onset of the illness, a significant increase in viral antibody titers was not observed. The patient had a slight fever with generalized maculopapular skin rash and an increased number of atypical lymphocytes, which is quite different from the typical clinical features of exanthem subitum.
Asunto(s)
Exantema Súbito/diagnóstico , Herpesvirus Humano 6 , Anticuerpos Antivirales/sangre , Exantema Súbito/inmunología , Exantema Súbito/virología , Femenino , Herpesvirus Humano 6/inmunología , Herpesvirus Humano 6/aislamiento & purificación , Humanos , Inmunidad Materno-Adquirida , Recién Nacido , Masculino , Embarazo , Viremia/inmunología , Viremia/virologíaRESUMEN
The reliability of loop-mediated isothermal amplification (LAMP), initially developed for the detection of human herpesvirus 7 (HHV-7), was evaluated in this study. Although a LAMP product was detected in HHV-7 DNA, neither HHV-6 nor human cytomegalovirus DNA produced a product. When agarose gel electrophoresis was used for the detection of LAMP products, the sensitivity of a 30-min HHV-7 LAMP reaction reached 250 copies/tube. The use of turbidity for the detection of the LAMP products gave a sensitivity of 500 and 250 copies/tube for 30- and 60-min reactions, respectively. Following these initial validation studies, clinical samples collected from two patients with primary HHV-7 infections were examined by HHV-7 LAMP. By use of agarose gel electrophoresis, HHV-7 LAMP products could be detected in acute-phase plasma samples but no LAMP product was detectable in convalescent-phase plasma samples from either patient. Since a turbidity assay is less sensitive than agarose gel electrophoresis, no HHV-7 LAMP product could be detected in plasma samples after a 30-min LAMP reaction. After a 60-min LAMP reaction, HHV-7 LAMP product could be detected in acute-phase plasma samples.
Asunto(s)
ADN Viral/genética , Herpesvirus Humano 7/genética , Herpesvirus Humano 7/aislamiento & purificación , Secuencia de Bases , Cartilla de ADN , ADN Viral/sangre , ADN Viral/química , Amplificación de Genes , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa/métodos , Infecciones por Roseolovirus/sangre , Infecciones por Roseolovirus/virologíaRESUMEN
A female infant developed Guillain-Barré syndrome 20 days after having exanthem subitum confirmed serologically as human herpesvirus 6 infection. DNA of human herpesvirus 6 was detected in peripheral blood mononuclear cells collected on admission.