RESUMEN
The use of improved microbiological procedures associated with molecular techniques has increased the identification of Candida bloodstream infections, even if the isolation of more than one species by culture methods remains uncommon. We report the cases of two children presenting with severe gastrointestinal disorders and other risk factors that contribute to Candida infections. In the first patient, C. albicans DNA was initially detected by a nested-amplification and C. tropicalis was found later during hospitalization, while blood cultures were persistently negative. In the second child, there was amplification of C. albicans and C. glabrata DNA in the same samples, but blood cultures yielded only C. albicans. Both patients received antifungal therapy but had unfavorable outcomes. These two cases illustrate that PCR was more successful than culture methods in detecting Candida in the bloodstream of high risk children, and was also able to detect the presence of more than one species in the same patient that might impact therapy when the fungi are resistant to azole compounds.
Asunto(s)
Candidemia/diagnóstico , Micología/métodos , Reacción en Cadena de la Polimerasa/métodos , Candida albicans/aislamiento & purificación , Candida glabrata/aislamiento & purificación , Candida tropicalis/aislamiento & purificación , Candidemia/microbiología , Niño , Preescolar , Enfermedad Crítica , Femenino , Enfermedades Gastrointestinales/complicaciones , Humanos , MasculinoRESUMEN
OBJECTIVE: To assess the testicular Sertoli cell function in male SLE patients. METHODS: Thirty-four consecutive patients were prospectively selected to evaluate serum inhibin B. Clinical features, treatment, semen analysis, urological evaluation, testicular ultrasound, hormones and anti-sperm antibodies were determined. RESULTS: Patients were subdivided into two groups: low serum inhibin B (Group 1, n = 8) and normal levels (Group 2, n = 26). The median sperm concentration (P = 0.024), total sperm count (P = 0.023) and total motile sperm count (P = 0.025) were lower in Group 1. Inhibin B levels were positively correlated with sperm concentration (r = 0.343), total motile sperm count (r = 0.357), and negatively correlated with follicule-stimulating hormone (FSH) (r = 0.699) and luteinizing hormone (r = 0.397). The median serum inhibin B was lower in SLE patients treated with intravenous cyclophosphamide (IVCYC) compared with those without this therapy (P = 0.031). Further evaluation of the 26 SLE patients with normal inhibin B and FSH levels revealed that medians of inhibin B/FSH ratio were lower in SLE patients with oligozoospermia compared with normozoospermia (P = 0.004). This ratio was also lower in SLE patients treated with IVCYC than those without this therapy (P = 0.04). In contrast, inhibin B serum level alone did not discriminate the later group of patients (P = 0.12). CONCLUSIONS: This is the first study to identify a high frequency of testicular Sertoli cell dysfunction in male SLE associated with semen abnormalities. Further prospective studies are necessary to determine if inhibin levels and inhibin B/FSH ratio will be an earlier and useful marker of IVCYC toxicity in these patients.
Asunto(s)
Lupus Eritematoso Sistémico/patología , Células de Sertoli/fisiología , Adolescente , Adulto , Autoanticuerpos/sangre , Distribución de Chi-Cuadrado , Ciclofosfamida/uso terapéutico , Hormona Folículo Estimulante/sangre , Humanos , Inmunosupresores/uso terapéutico , Inhibinas/sangre , Lupus Eritematoso Sistémico/sangre , Lupus Eritematoso Sistémico/tratamiento farmacológico , Hormona Luteinizante/sangre , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Células de Sertoli/metabolismo , Células de Sertoli/patología , Recuento de Espermatozoides , Motilidad Espermática , Espermatozoides/inmunología , Estadísticas no Paramétricas , Testículo/diagnóstico por imagen , UltrasonografíaRESUMEN
A rapid DNA extraction was used for T. cruzi detection in triatomines dry fecal spots collected on filter paper and analyzed by PCR. Fifty T. infestans were fed on experimentally infected Balb/C mice with high T. cruzi parasitemia and divided into five groups of ten triatomines, and 100 triatomines were infected with lower parasitemia and divided into five groups of 20 triatomines. One dry fecal spot was analyzed per group on days 1, 2, 3, 4 and 5 post feeding. Amplification targeted T. cruzi TCZ sequence and resulted positive from day 4 after bugs feeding in the two models (high and lower parasitemial. The rapid DNA isolation and PCR proposed are suitable for detection of T. cruzi DNA in filter paper and should be considered in field research.
Asunto(s)
Insectos Vectores/parasitología , Triatoma/parasitología , Trypanosoma cruzi/aislamiento & purificación , Animales , Enfermedad de Chagas/diagnóstico , Enfermedad de Chagas/parasitología , ADN Protozoario/química , ADN Protozoario/genética , Heces/parasitología , Amplificación de Genes , Humanos , Parasitemia/diagnóstico , Parasitemia/parasitología , Reacción en Cadena de la Polimerasa/métodos , Sensibilidad y Especificidad , Trypanosoma cruzi/genéticaAsunto(s)
Bartonella henselae/aislamiento & purificación , Donantes de Sangre , Portador Sano/sangre , Enfermedad por Rasguño de Gato/sangre , Adulto , Animales , Bartonella henselae/fisiología , Bartonella henselae/ultraestructura , Portador Sano/microbiología , Enfermedad por Rasguño de Gato/microbiología , Gatos , Eritrocitos/microbiología , Eritrocitos/ultraestructura , Reacciones Falso Negativas , Femenino , Técnica del Anticuerpo Fluorescente Indirecta , Humanos , Lactante , Microscopía Electrónica , Reacción en Cadena de la Polimerasa/métodosRESUMEN
In order to study B. henselae transmission among cats, five young cats were kept in confinement for two years, one of them being inoculated by SC route with B. henselae (10(5) UFC). Only occasional contact among cats occurred but the presence of fleas was observed in all animals throughout the period. Blood culture for isolation of bacteria, PCR-HSP and FTSZ (gender specific), and BH-PCR (species-specific), as well as indirect immunofluorescence method for anti-B. henselae antibodies were performed to confirm the infection of the inoculated cat as well as the other naive cats. Considering the inoculated animal, B. henselae was first isolated by blood culture two months after inoculation, bacteremia last for four months, the specific antibodies being detected by IFI during the entire period. All contacting animals presented with bacteremia 6 months after experimental inoculation but IFI did not detect seroconversion in these animals. All the isolates from these cats were characterized as Bartonella (HSP and FTSZ-PCR), henselae (BH-PCR). However, DNA of B. henselae could not be amplified directly from peripheral blood by the PCR protocols used. Isolation of bacteria by blood culture was the most efficient method to diagnose infection compared to PCR or IFI. The role of fleas in the epidemiology of B. henselae infection in cats is discussed.
Asunto(s)
Anticuerpos Antibacterianos/análisis , Infecciones por Bartonella/veterinaria , Bartonella henselae/inmunología , Enfermedades de los Gatos/transmisión , Transmisión de Enfermedad Infecciosa/veterinaria , Animales , Infecciones por Bartonella/diagnóstico , Infecciones por Bartonella/transmisión , Enfermedades de los Gatos/diagnóstico , Gatos , ADN Bacteriano/análisis , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Reacción en Cadena de la Polimerasa , Sensibilidad y EspecificidadRESUMEN
We present a case of prenatal diagnosis of congenital rubella. After birth, in addition to traditional serologic and clinical examinations to confirm the infection, we could identify the virus in the "first fluid aspirated from the oropharynx of the newborn", using polimerase chain reaction (PCR). We propose that this first oropharynx fluid (collected routinely immediately after birth) could be used as a source for identification of various congenital infection agents, which may not always be easily identified by current methods.
Asunto(s)
Orofaringe/virología , Reacción en Cadena de la Polimerasa , Síndrome de Rubéola Congénita/diagnóstico , Adulto , Femenino , Humanos , Recién Nacido , Masculino , Orofaringe/metabolismo , Embarazo , Complicaciones Infecciosas del Embarazo/diagnóstico , Diagnóstico Prenatal , ARN Viral/aislamiento & purificaciónAsunto(s)
Diabetes Mellitus Tipo 2/complicaciones , Lupus Eritematoso Sistémico/complicaciones , Síndrome de Necrosis Retiniana Aguda/parasitología , Toxoplasmosis Ocular/complicaciones , Femenino , Angiografía con Fluoresceína , Humanos , Persona de Mediana Edad , Síndrome de Necrosis Retiniana Aguda/patologíaAsunto(s)
ADN Viral/análisis , Infecciones por VIH/diagnóstico , VIH-1 , Humanos , Lactante , ARN Viral/análisisRESUMEN
Rubella infection is an exanthematic disease, with high prevalence in the adult population. The only modality of disease that causes serious consequences is congenital rubella syndrome (CRS), which happens when a pregnant woman seronegative to rubella virus acquires the infection during early pregnancy. Due to the lack of signals and characteristic symptoms of disease, diagnosis of rubella is based essentially on laboratory tests: antibodies detection and/or virus isolation. Results of serologic tests should always be interpreted with caution, because they can be affected by the quality of blood samples, processing and storage of sera, the equipment and reagents used to perform tests, and finally by the technical expertise and training of biologists. The collection tubes with gel seem to facilitate serum separation, but on the other hand gels can retain and consequently decrease antibody titers. Therefore, we decided to investigate whether the use of collection tubes containing gel separator might interfere with rubella virus antibody detection in blood samples from children. We did not observe statistically significant differences with respect to rubella virus antibody detection (immunoglobulin M [IgM] and immunoglobulin G [IgG]) for samples collected in tubes with or without gel separator, from the two evaluated manufacturers.
Asunto(s)
Anticuerpos Antivirales/sangre , Recolección de Muestras de Sangre/instrumentación , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Niño , Geles , HumanosRESUMEN
The only threatening modality of rubella is the Congenital Rubella Syndrome that affects fetuses of women who acquire infection during early pregnancy. Laboratory diagnosis is based on serological parameters. We compared anti-rubella IgM and IgG detection of two commercial immunoassay kits (Abbott and Roche). Although we observed an agreement of 97.8% for IgM and 95.7% for IgG when the categories positive, negative and indeterminate were considered, mean titers of IgG and the absorbance/cut off of IgM were statistically different for both kits, thus corroborating the idea that serological results depend very much on the methodology and must be carefully interpreted.
Asunto(s)
Anticuerpos Antivirales/sangre , Inmunoensayo/métodos , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Juego de Reactivos para Diagnóstico , Rubéola (Sarampión Alemán)/diagnóstico , Adolescente , Anticuerpos Antivirales/análisis , Niño , Preescolar , Humanos , Inmunoglobulina G/análisis , Inmunoglobulina M/análisis , Rubéola (Sarampión Alemán)/sangre , Rubéola (Sarampión Alemán)/inmunologíaRESUMEN
AIMS: To evaluate whether procalcitonin (PCT) and C reactive protein (CRP) are able to discriminate between sepsis and systemic inflammatory response syndrome (SIRS) in critically ill children. METHODS: Prospective, observational study in a paediatric intensive care unit. Kinetics of PCT and CRP were studied in patients undergoing open heart surgery with cardiopulmonary bypass (CPB) (SIRS model; group I1) and patients with confirmed bacterial sepsis (group II). RESULTS: In group I, PCT median concentration was 0.24 ng/ml (reference value <2.0 ng/ml). There was an increment of PCT concentrations which peaked immediately after CPB (median 0.58 ng/ml), then decreased to 0.47 ng/ml at 24 h; 0.33 ng/ml at 48 h, and 0.22 ng/ml at 72 h. CRP median concentrations remained high on POD1 (36.6 mg/l) and POD2 (13.0 mg/l). In group II, PCT concentrations were high at admission (median 9.15 ng/ml) and subsequently decreased in 11/14 patients who progressed favourably (median 0.31 ng/ml). CRP levels were high in only 11/14 patients at admission. CRP remained high in 13/14 patients at 24 h; in 12/14 at 48 h; and in 10/14 patients at 72 h. Median values were 95.0, 50.9, 86.0, and 20.3 mg/l, respectively. The area under the ROC curve was 0.99 for PCT and 0.54 for CRP. Cut off concentrations to differentiate SIRS from sepsis were >2 ng/ml for PCT and >79 mg/l for CRP. CONCLUSION: PCT is able to differentiate between SIRS and sepsis while CRP is not. Moreover, unlike CRP, PCT concentrations varied with the evolution of sepsis.
Asunto(s)
Infecciones Bacterianas/diagnóstico , Proteína C-Reactiva/análisis , Calcitonina/sangre , Complicaciones Posoperatorias/diagnóstico , Precursores de Proteínas/sangre , Sepsis/diagnóstico , Síndrome de Respuesta Inflamatoria Sistémica/diagnóstico , Adolescente , Biomarcadores/sangre , Proteína C-Reactiva/metabolismo , Péptido Relacionado con Gen de Calcitonina , Puente Cardiopulmonar , Niño , Preescolar , Diagnóstico Diferencial , Femenino , Cardiopatías Congénitas/cirugía , Humanos , Lactante , Recién Nacido , Masculino , Estudios ProspectivosRESUMEN
UNLABELLED: A prospective study was conducted to determine if standardized vancomycin doses could produce adequate serum concentrations in 25 term newborn infants with sepsis. PURPOSE: The therapeutic response of neonatal sepsis by Staphylococcus sp. treated with vancomycin was evaluated through serum concentrations of vancomycin, serum bactericidal titers (SBT), and minimum inhibitory concentration (MIC). METHOD: Vancomycin serum concentrations were determined by the fluorescence polarization immunoassay technique, SBT by the macro-broth dilution method, and MIC by diffusion test in agar. RESULTS: Thirteen newborn infants (59.1%) had adequate peak vancomycin serum concentrations (20 - 40 mg/mL) and one had peak concentration with potential ototoxicity risk (>40 microg/mL). Only 48% had adequate trough concentrations (5 - 10 mg/mL), and seven (28%) had a potential nephrotoxicity risk (>10 microg/mL). There was no significant agreement regarding normality for peak and trough vancomycin method (McNemar test : p = 0.7905). Peak serum vancomycin concentrations were compared with the clinical evaluation (good or bad clinical evolution) of the infants, with no significant difference found (U=51.5; p=0.1947). There was also no significant difference between the patients' trough concentrations and good or bad clinical evolution (U = 77.0; p=0.1710). All Staphylococcus isolates were sensitive to vancomycin according to the MIC. Half of the patients with adequate trough SBT (1/8), also had adequate trough vancomycin concentrations and satisfactory clinical evolution. CONCLUSIONS: Recommended vancomycin schedules for term newborn infants with neonatal sepsis should be based on the weight and postconceptual age only to start antimicrobial therapy. There is no ideal pattern of vancomycin dosing; vancomycin dosages must be individualized. SBT interpretation should be made in conjunction with the patient's clinical presentation and vancomycin serum concentrations. Those laboratory and clinical data favor elucidation of the probable cause of patient's bad evolution, which would facilitate drug adjustment and reduce the risk of toxicity or failing to achieve therapeutic doses.
Asunto(s)
Antibacterianos/administración & dosificación , Sepsis/tratamiento farmacológico , Infecciones Estafilocócicas/tratamiento farmacológico , Vancomicina/administración & dosificación , Esquema de Medicación , Humanos , Recién Nacido , Pruebas de Sensibilidad Microbiana , Estudios Prospectivos , Prueba Bactericida de Suero , Estadísticas no ParamétricasRESUMEN
INTRODUCTION: Peak and trough serum concentrations of vancomycin were determined in term newborn infants with confirmed or suspected Staphylococcus sp sepsis by high performance liquid chromatography and flourescence polarization immunoassay. OBJECTIVE: To statistically compare the results of the high performance liquid chromatography and flourescence polarization immunoassay techniques for measuring serum vancomycin concentrations. METHODS: Eighteen peak and 20 trough serum samples were assayed for vancomycin concentrations using high performance liquid chromatography and flourescence polarization immunoassay from October 1995 to October 1997. RESULTS: The linear correlation coefficients for high performance liquid chromatography and flourescence polarization immunoassay were 0.27 (peak, P = 0.110) and 0.26 (trough, P = 0.1045) respectively, which were not statistically significant. CONCLUSION: There was wide variation in serum vancomycin concentrations determined by high performance liquid chromatography as compared with those determined by flourescence polarization immunoassay. There was no recognizable pattern in the variability; in an apparently random fashion, the high performance liquid chromatography measurement was sometimes substantially higher than the flourescence polarization immunoassay measurement, and at other times it was substantially lower.
Asunto(s)
Antibacterianos/sangre , Sepsis/sangre , Infecciones Estafilocócicas/sangre , Vancomicina/sangre , Cromatografía Líquida de Alta Presión , Inmunoensayo de Polarización Fluorescente , Humanos , Recién Nacido , Monitoreo Fisiológico , Sepsis/tratamiento farmacológico , Infecciones Estafilocócicas/tratamiento farmacológicoAsunto(s)
Enfermedad de Chagas/diagnóstico , Insectos Vectores/parasitología , Triatoma/parasitología , Trypanosoma cruzi/aislamiento & purificación , Animales , Enfermedad de Chagas/genética , Enfermedad de Chagas/parasitología , Cartilla de ADN/genética , ADN Protozoario/genética , Humanos , Ratones , Ratones Endogámicos BALB C , Microscopía/métodos , Reacción en Cadena de la Polimerasa/métodos , Secuencias Repetitivas de Ácidos Nucleicos/genética , Trypanosoma cruzi/genéticaRESUMEN
Relatamos um caso de diagnostico pre-natal de rubeola congenita. Apos o nascimento, alem da confirmacao feita atraves do exame fisico e sorologico do recem-nascido, o virus tambem pode ser demonstrado no primeiro fluido aspirado da orofaringe do recem-nascido, utilizando-se a reacao em cadeia da polimerase (PCR). Sugerimos que este fluido (colhido rotineiramente no momento da reanimacao neonatal) possa ser utilizado na pesquisa de outros agentes infecciosos, que nao sao facilmente identificados por outros metodos