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BACKGROUND: Oronasal fistula is one of the most critical complications in primary palatoplasty, with a reported incidence between 5% and 46% with multiple associated risk factors described previously. In addition, in more than half of the patients, it implies additional surgeries, increasing risks for the patient and health care costs. In this case-control study, the authors aim to determine the specific risk factors for oronasal fistula after primary palatoplasty in the study population. METHODS: A retrospective review was undertaken to identify all patients undergoing primary palatoplasty between 2017 and 2019. Patients who developed oronasal fistula were included (cases) and compared with patients from the same cohort without fistula (controls). Demographic, clinical, and perioperative variables and their association with postoperative fistula presentation were explored through crude and adjusted analysis. RESULTS: One hundred thirty-nine patients with a median age of 6 years (5-6) who underwent primary palatoplasty were found. Forty-five presented an oronasal fistula, corresponding to 32% of the population, and 64% required subsequent repair of an oronasal fistula. In the crude analysis, the surgeon's experience (OR: 0.44) was associated significantly with the outcome. Adjusted analysis showed an association between surgical site infection and syndromic presentation. CONCLUSIONS: The experience of the surgeon is a protective factor for the oronasal fistula presentation; in addition, the presentation of infection of the operative site, the syndromic presentation, and the clinical follow-up at 1 year were also relevant; the latter, possibly due to the social context of our patients.
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Resumen Introducción: anualmente más de 180.000 personas mueren por quemaduras. Existen varios índices para evaluar la mortalidad en pacientes quemados. El índice de área letal 50 (AL50) permite observar y comparar resultados entre diferentes unidades de quemados. Objetivo: determinar el AL50 en la unidad de quemados de un hospital de referencia en Medellín, Colombia entre 2015 y 2019, identificar la tendencia en los últimos años y compararla con la reportada por otros centros de atención. Materiales y métodos: se hizo un estudio retrospectivo. Se incluyeron pacientes con quemaduras mayores al 19 % de superficie corporal total quemada que ingresaron al Hospital San Vicente Fundación entre enero del 2015 y diciembre del 2019. Se analizaron los resultados de forma global y por subgrupos de edad. Se realizó un modelo probit para el cálculo del área letal 50. Resultados: se admitieron 590 pacientes en el análisis, con una media de edad de 30 años, 69,6 % hombres y 30,4 % mujeres. La media de superficie corporal total quemada fue 33 %, el mecanismo de lesión más común fue por llama con un 55 % y el AL50 encontrado fue del 68 %. En el análisis por subgrupos se encontró un AL50 de 77,8 % en pacientes entre 0 y 14 años, 71,8 % entre 15 y 44 años, 60,4 % entre 45 y 64 años y 53,1 % para mayores de 65 años. Conclusiones: el AL50 calculado fue de 68 %, comparable al encontrado en otros centros de referencia a nivel internacional. Este varía según los rangos de edad, y no hubo tendencia al aumento con el paso de los años.
Abstract Introduction: Burns result in more than 180,000 deaths annually. Various indices exist to assess burn-related mortality. The lethal area 50 (LA50) index enables the evaluation and comparison of outcomes among different burn units. Objective: This study aimed to determine the LA50 within the burn unit of a reference hospital in Medellín, Colombia, between 2015 and 2019. It also aimed to identify trends over recent years and compare the findings with those reported by other healthcare facilities. Materials and Methods: A retrospective study was conducted including patients admitted to Hospital San Vicente Fundación between January 2015 and December 2019. Participants with burns exceeding 19% of the total burned body surface area were included. Data were analyzed both globally and in age subgroups. A probit model was used to calculate the LA50. Results: A total of 590 patients were included in the analysis, with a mean age of 30 years. Of these, 69.6% were male and 30.4% were female. The mean total body surface area burned was 33%, with flame being the most common injury mechanism (55%). The calculated LA50 was 68%. Subgroup analysis revealed LA50 values of 77.8% for patients aged 0 to 14 years, 71.8% for those aged 15 to 44 years, 60.4% for individuals aged 45 to 64 years, and 53.1% for patients over 65 years. Conclusions: The calculated LA50 of 68% is comparable to that reported by other internationally recognized centers. The LA50 varies according to age ranges, and there was no increasing trend observed over the years.
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Vaginal infection is a gynecological problem in women of reproductive age with multiple health outcomes. The most common forms of infection include bacterial vaginosis (BV), vulvovaginal candidiasis (VC), and aerobic vaginitis (AV). Our main goals were to evaluate different types of vaginal infections in Ecuadorian women in a large urban area (Quito) and to characterize the vaginal microbiota colonization by opportunistic species. We collected vaginal swabs and epidemiological surveys from 414 women from June 2016 to July of 2017. We analyzed vaginal samples for the presence of any vaginal infection. The microbiological examination was done through Gram-stain, wet mount smears, and polymerase chain reaction (PCR) assays using primers for target genes, such as 16S rRNA (Atopobium vaginae, Mobiluncus mulieris, and Gardnerella species), ddl (Enterococcus faecalis), adk (Escherichia coli) and KER1 (Candida albicans) genes. Most women showed a healthy vaginal microbiota (66.7%). Nearly one-tenth (10.4%) of the participants had intermediate microbiota, and the remaining women (22.9%) had a single vaginal infection (BV, AV, or VC) or coinfections. From the 95 participants that had an infection, AV was the main diagnosed vaginal infection (51.6%), followed by BV (24.2%) and finally VC (7.4%). The remaining women (16.8%) showed coinfections, being BV and AV the most common coinfection. Using univariable logistic regression analyses we found an increased odds of healthy microbiota in women with a sexual partner (P = 0.02, OR = 1.64). Also, women in a free union relationship (P = 0.000, OR = 16.65) had an increased odds of having coinfections. On the other hand, the use of birth control (condom OR = 0.388 or other contraceptive method OR = 0.363) was associated with significantly lower odds of intermediate microbiota (P ≤ 0.05). We found no statistically significant differences between women with infection and a particular group age. Using multivariate logistic regression analyses we initially found an increased odds of having BV in women with M. mulieris (P = 0.020, OR = 4.98) and Gardnerella species (P = 0.010, OR = 4.16). Women with E. coli showed an increased odds of having AV (P = 0.009, OR = 2.81). The presence of C. albicans in women showed an increased odds of having VC (P = 0.007, OR = 17.94). Finally, women with M. mulieris showed a reverse odds of having healthy microbiota (P = 0.008, OR = 0.06). We found no statistically significant differences between women with symptomatic and asymptomatic infections or the presence of Enterococcus faecalis. We found using logistic regression analyses that M. mulieris was the most prevalent opportunistic pathogen among women with vaginal infection. Further studies should evaluate the possibility to use M. mulieris as a potential key predictor for vaginal infections.
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Microbiota , Vagina/microbiología , Enfermedades Vaginales/microbiología , Adulto , Factores de Edad , Candidiasis Vulvovaginal/epidemiología , Candidiasis Vulvovaginal/microbiología , Ecuador/epidemiología , Femenino , Humanos , Persona de Mediana Edad , Prevalencia , Factores de Riesgo , Enfermedades Vaginales/epidemiología , Vaginosis Bacteriana/epidemiología , Vaginosis Bacteriana/microbiología , Adulto JovenRESUMEN
ABSTRACT Cutaneous mucinosis is a group of conditions characterized by the abnormal deposition of mucin in the skin. They can be primary, which in turn can be inflammatory-degenerative, and hamartomatous-neoplastic; or secondary. Papulonodular mucinosis is part of the group of dermal inflammatory-degenerative primary mucinosis. Its association with autoimmune connective tissue diseases has been described, especially with systemic lupus erythematosus, but it is considered an unusual manifestation of this disease. The clinical case is presented of an 11 year-old girl who, at the onset of systemic lupus erythematosus, presented with skin lesions for which the histopathological diagnosis corresponded to mucinosis.
RESUMEN Las mucinosis cutáneas son un grupo de condiciones caracterizadas por el depósito anormal de mucina en la piel. Pueden ser primarias, que a su vez pueden ser inflamatorias-degenerativas (dérmicas o foliculares) y hamartomatosas-neoplásicas; o secundarias. La mucinosis papulonodular forma parte de las mucinosis primarias inflamatorias-degenerativas dérmicas. Se ha descrito su asociación con enfermedades autoinmunes del tejido conectivo, especialmente con el lupus eritematoso sistémico, pero se considera una manifestación inusual de esta enfermedad. Se presenta el caso clínico de una niña de 11 años, quien al inicio del lupus eritematoso sistémico presentaba lesiones en la piel cuyo diagnóstico histopatológico correspondió a mucinosis.
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Humanos , Femenino , Niño , Mucinosis , Lupus Eritematoso Sistémico , Asociación , Piel , Heridas y LesionesRESUMEN
BACKGROUND: Bacterial vaginosis (BV) is a microbial imbalance (i.e., dysbiosis) that can produce serious medical effects in women at childbearing age. Little is known, however, about the incidence of BV or vaginal microbiota dysbiosis in pregnant teenagers in low and middle-income countries such as Ecuador. The scope of this exploratory analysis was to study the relationship between epidemiologic and microbial risk factors. Among the microbiology risk factors this study investigated five Lactobacillus species, two of them know in preview studies as microbiology risk factors for BV development (Lactobacillus acidophilus and Lactobacillus iners), and the last three known for being associated with a healthy vaginal tract (Lactobacillus crispatus, Lactobacillus gasseri and Lactobacillus jensenii). In addition, fastidious anaerobes known to be microbial risk factors for BV development in pregnant teenagers were searched as well, more exactly, Gardnerella vaginalis, Atopobium vaginae and Mobiluncus mulieris. METHODS: Ninety-five healthy adolescent pregnant women, visiting a secondary level hospital in Quito, Ecuador, were enrolled into the study in 2015. The enrolled patients were between 10 to 13 weeks of pregnancy. Four epidemiological risk factors were collected in a survey: age, civil status, sexual partners and condom use. Also, vaginal pH was measured as a health risk factor. DNA was extracted from endocervical and exocervical epithelia from all the patients' samples. PCR analysis was performed in order to characterize the presence of the eight bacterial species known as risk factors for BV development, targeting three anaerobes and five Lactobacillus species. Univariate and multivariate analysis were performed to identify associated factors for the presence of anaerobic species using logistic regression. RESULTS: The 95 vaginal microflora samples of these teenagers were analyzed. Two of the bacterial species known to cause BV: A. vaginae (100%) and G. vaginalis (93.7%) were found in high prevalence. Moreover, the most predominant bacterial Lactobacillus species found in the pregnant teenagers' vaginal tract were L. crispatus (92.6%), L. iners (89.5%) and L. acidophilus (87.4%). In addition, the average vaginal pH measured in the study population was 5.2, and high pH was associated with the presence of the three-anaerobic species (p = 0.001). Finally, L. jensenii's presence in the study decreased in 72% the occupation of the three anaerobes. DISCUSSION: This work identified a high pH as a risk factor for BV anaerobes' presence in adolescent pregnant women. Moreover, this study identified L. crispatus, L. iners and L. acidophilus to be the most abundant species in our study population. From all fastidious anaerobes analyzed in this study, A. vaginae was present in all pregnant teenagers. To conclude, L. jensenii could be a potential healthy vaginal microbiota candidate in pregnant teenagers and should be further analyzed in future studies.
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El síndrome de hipoventilación central congénito idiopático, también conocido como síndrome de Ondina, es una enfermedad poco común, caracterizada por la ausencia congénita del control central de la respiración y disfunción del sistema nervioso autónomo. Su incidencia se estima en aproximadamente 1 de cada 200 000 nacimientos al año. Es una enfermedad de transmisión autosómica dominante, derivada de una mutación heterocigótica del gen PHOX2B, presente en 90 % de los pacientes, pero su causa fisiopatológica aún no está claramente dilucidada. Presenta una elevada tasa de mortalidad, y una dependencia a la ventilación mecánica durante el sueño de por vida; sin embargo, gracias a una atención multidisciplinar y coordinada, con estrecha vigilancia y apoyo, podría esperarse que los pacientes con ella puedan llevar una vida relativamente normal. Se realiza el reporte de un caso diagnosticado en el departamento de Antioquia (Colombia), lugar con importantes barreras socioeconómicas que limitan el estudio complejo de este tipo de enfermedades de baja prevalencia global.
Idiopathic congenital central hypoventilation syndrome, also known as Ondine´s curse, is a rare disease characterized by congenital absence of the central control of breathing and autonomic nervous system dysfunction. Its incidence is estimated to be one per 200,000 births a year. It is an autosomal dominant disease derived from a heterozygous PHOX2B gene mutation, present in 90 % of patients, but its pathophysiological cause has not been elucidated yet. It has a high mortality rate and a lifelong dependency on a life-support device during sleep. However, it could be expected that a coordinated multidisciplinary care, with close monitoring and support, could help this kind of patients to have a relatively normal life. This is the report of a case diagnosed with this disease in Antioquia, Colombia, a place with significant socio-economic limitations that hinder a complete study of this type of low overall prevalence disease.
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BACKGROUND: Helicobacter pylori, the main cause of peptic ulcer disease and gastric cancer in adult populations, is generally acquired during the first years of life. Infection can be persistent or transient and bacterial and host factors determining persistence are largely unknown and may prove relevant for future disease. METHODS: Two cohorts of healthy Chilean infants (313 total) were evaluated every 3 months for 18-57 months to determine pathogen- and host-factors associated with persistent and transient infection. RESULTS: One-third had at least one positive stool ELISA by age 3, with 20% overall persistence. Persistent infections were acquired at an earlier age, associated with more household members, decreased duration of breastfeeding, and nonsecretor status compared to transient infections. The cagA positive strains were more common in persistent stools, and nearly 60% of fully characterized persistent stool samples amplified cagA/vacAs1m1. Persistent children were more likely to elicit a serologic immune response, and both infection groups had differential gene expression profiles, including genes associated with cancer suppression when compared to healthy controls. CONCLUSIONS: These results indicate that persistent H. pylori infections acquired early in life are associated with specific host and/or strain profiles possibly associated with future disease occurrence.
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Heces/microbiología , Infecciones por Helicobacter/epidemiología , Helicobacter pylori/aislamiento & purificación , Anticuerpos Antibacterianos/sangre , Antígenos Bacterianos/genética , Enfermedades Asintomáticas , Proteínas Bacterianas/genética , Preescolar , Chile/epidemiología , Estudios de Cohortes , Ensayo de Inmunoadsorción Enzimática , Perfilación de la Expresión Génica , Infecciones por Helicobacter/inmunología , Infecciones por Helicobacter/microbiología , Helicobacter pylori/genética , Helicobacter pylori/patogenicidad , Interacciones Huésped-Patógeno , Humanos , Lactante , Reacción en Cadena de la Polimerasa , Estudios Prospectivos , Factores de TiempoRESUMEN
BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a disorder associated to cigarette smoke and lung cancer (LC). Since epigenetic changes in oncogenes and tumor suppressor genes (TSGs) are clearly important in the development of LC. In this study, we hypothesize that tobacco smokers are susceptible for methylation in the promoter region of TSGs in airway epithelial cells when compared with non-smoker subjects. The purpose of this study was to investigate the usefulness of detection of genes promoter methylation in sputum specimens, as a complementary tool to identify LC biomarkers among smokers with early COPD. METHODS: We determined the amount of DNA in induced sputum from patients with COPD (n = 23), LC (n = 26), as well as in healthy subjects (CTR) (n = 33), using a commercial kit for DNA purification, followed by absorbance measurement at 260 nm. The frequency of CDKN2A, CDH1 and MGMT promoter methylation in the same groups was determined by methylation-specific polymerase chain reaction (MSP). The Fisher's exact test was employed to compare frequency of results between different groups. RESULTS: DNA concentration was 7.4 and 5.8 times higher in LC and COPD compared to the (CTR) (p < 0.0001), respectively. Methylation status of CDKN2A and MGMT was significantly higher in COPD and LC patients compared with CTR group (p < 0.0001). Frequency of CDH1 methylation only showed a statistically significant difference between LC patients and CTR group (p < 0.05). CONCLUSIONS: We provide evidence that aberrant methylation of TSGs in samples of induced sputum is a useful tool for early diagnostic of lung diseases (LC and COPD) in smoker subjects. VIRTUAL SLIDES: The abstract MUST finish with the following text: Virtual Slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1127865005664160.
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ADN/análisis , Detección Precoz del Cáncer/métodos , Genes Supresores de Tumor , Neoplasias Pulmonares/diagnóstico , Enfermedad Pulmonar Obstructiva Crónica/diagnóstico , Esputo/química , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/genética , Metilación de ADN/genética , Humanos , Neoplasias Pulmonares/genética , Persona de Mediana Edad , Regiones Promotoras Genéticas/genética , Enfermedad Pulmonar Obstructiva Crónica/genética , Fumar/efectos adversos , Adulto JovenAsunto(s)
Humanos , Femenino , Médicos Mujeres , Médicos Mujeres/historia , Rol del Médico , Mujeres TrabajadorasRESUMEN
La infertilidad asociada a trastornos en la espermatogénesis y en la calidad seminal, es un problema de salud pública cuyo manejo terapéutico actual produce efectos colaterales, carece de eficacia significativa y el paciente habitualmente termina en procedimientos de reproducción asistida. Actualmente una alternativa plausible para enfrentar estos problemas es el uso de plantas medicinales; tal es el caso de Lepidium meyenii (Maca). En el presente trabajo se estudió el efecto de una variedad local de Maca sobre la espermatogénesis y la calidad espermática en diez sujetos diagnosticados de infertilidad y que presentaban alteraciones en uno o más parámetros seminales. El tratamiento con Lepidium meyenii (Maca) duró 3 meses, en una dosis de 3000 mg/día; se evaluaron todos los parámetros seminales, de acuerdo a las guías de la OMS y se determinaron las concentraciones de Testosterona, Hormona Folículo Estimulante (FSH), Estradiol, Sulfato de Dehidroepiandrosterona (DHEA-S), Hormona Estimulante de la Tiroides (TSH) y de Antígeno Prostático Específico (PSA) séricas antes y después del tratamiento. La motilidad espermática Grado III fue significativamente mejorada: en la 1ª hora antes del tratamiento presentó valores de 10 ± 0.06, en relación a 15 ± 0.07 después del tratamiento (P < 0.001), en la 3ª hora los valores de 8 ± 0.06 antes del tratamiento se incrementaron a 11 ± 0.06 (P < 0.001). Los valores de la motilidad progresiva (motilidad espermática grado III y II) a la 1ª hora antes del tratamiento, fueron de 67.2 ± 0.09, con aumento después del tratamiento a 74.3 ± 0.07 (P < 0.01). La vitalidad espermática en la 1ª hora no presentó cambios significativos con el tratamiento, en cambio a la 3ª hora sus valores antes del tratamiento fueron de 75 ± 0.08, y después del tratamiento fueron de 79 ± 0.07 (P < 0.01). La morfología presentó valores de espermatozoides normales antes del tratamiento de 40 ± 0.10 y después del tratamiento 45 ± 0.11 (P < 0.01). El porcentaje de células germinales o inmaduras antes del tratamiento fue de 5.6 ± 0.045 y después del tratamiento 3.7 ± 0.037 (P < 0.01). Las concentraciones hormonales antes y después del tratamiento no variaron significativamente. El Antígeno Prostático Especifico tampoco se modificó después del tratamiento. El efecto más evidente fue el incremento de la proporción de espermatozoides motiles en sujetos cuya deficiencia se expresa en este parámetro; este incremento correlacionó directamente con el hecho de que tales sujetos pudieron embarazar a sus parejas durante el tratamiento. (AU)
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Humanos , Masculino , Femenino , Espermatogénesis , Lepidium , Bolivia , Infertilidad , Medicina TradicionalAsunto(s)
Sedimentación Sanguínea , Patógenos Transmitidos por la Sangre , Contaminación de Equipos , Infección de Laboratorio/epidemiología , Ochrobactrum anthropi/aislamiento & purificación , Adolescente , Adulto , Anciano , Bacteriemia/epidemiología , Niño , Preescolar , Chile , Técnicas de Laboratorio Clínico , Brotes de Enfermedades , Femenino , Hospitales de Enseñanza , Humanos , Infección de Laboratorio/microbiología , Masculino , Persona de Mediana EdadRESUMEN
Coagulase-negative staphylococci (CNS) are frequently isolated from blood cultures, where they may be only a contaminant or the cause of bacteraemia. Determining whether an isolate of CNS represents a true CNS bacteraemia is difficult, and there is no single criterion with sufficient specificity. The aim of this study was to assess those clinical, microbiological, pathogenic and genotypic features that characterize true CNS bacteraemia. Twenty patients having two or more blood cultures positive for CNS and 20 patients with only one positive blood culture were studied. Significant bacteraemia was defined according to clinical and laboratory criteria. Incubation time for blood cultures to become positive, macroscopic appearance of colonies, species determination, biotype, susceptibility to antimicrobials, PFGE pattern and adherence capacity were all studied. Clinical bacteraemia was present in 16/20 patients with two or more positive blood cultures and in 2/20 patients with only one positive blood culture. A significant difference was seen in the median time to positivity between the 18 clinical bacteraemias and 22 contaminations (23.6 versus 29.2 h; P = 0.04, Wilcoxon). There was also a significant difference between the two groups in the median absorbance of the slime test (1.36 versus 0.58; P = 0.005). All significant bacteraemias with two or more positive blood cultures had the same species identified, the same antimicrobial susceptibility pattern and the same PFGE pattern. In two patients with true bacteraemia with only one positive blood culture, the incubation time for the culture to turn positive was <24 h and the slime production absorbance was >2.5. The most useful parameters for the diagnosis of true CNS bacteraemia for patients with two positive blood cultures were incubation time until positive, species identification, antimicrobial susceptibility pattern, slime production and PFGE pattern. For patients with only one blood culture positive for CNS, the useful parameters for prediction of true bacteraemia were incubation time until positive and slime production, both of which are simple, low-cost tests.
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Bacteriemia/microbiología , Infecciones Estafilocócicas/microbiología , Staphylococcus/patogenicidad , Adhesión Bacteriana , Coagulasa , Electroforesis en Gel de Campo Pulsado , Genotipo , Humanos , Pruebas de Sensibilidad Microbiana , Reacción en Cadena de la Polimerasa , Sensibilidad y Especificidad , Especificidad de la Especie , Staphylococcus/clasificación , Staphylococcus/enzimología , Staphylococcus/genética , VirulenciaRESUMEN
Un número importante de inmigrantes provenientes de áreas endémicas para enfermedad de Chagas viven en la VII región. Dado que una de las formas de transmisión del Chagas es por transfusión sanguínea, es importante evaluar, en zonas no endémicas, la utilidad de una encuesta de predonación para identificar donantes de sangre infectados con trypanosoma cruzi. En este trabajo participaron 7 hospitales de la región, durante un período de 6 meses, 1.581 donantes de sangre fueron encuestados y sus sueros analizados para detectar la presencia de anticuerpos anti- T. cruzi. La efectividad del cuestionario empleado fue evaluada, correlacionando las respuestas a las diferentes preguntas sobre la enfermedad, con el resultado del ensayo inmunoabsorberte ligado a enzima. Las muestras positivas fueron confirmadas por: inmunofluorescencia indirecta y reacción en cadena de la polimerasa. Solamente un donante fue positivo para enfermedad de Chagas. Este dador no reportó factores de riesgo. Po lo que donantes de sangre seropositivos y parasitémicos para T. cruzi, están presente en la población de donantes de sangre de zonas no endémicas, sin tener los usuales factores de riesgo
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Humanos , Donantes de Sangre/estadística & datos numéricos , Encuestas Epidemiológicas , Trypanosoma cruzi/aislamiento & purificación , Enfermedad de Chagas/prevención & control , Enfermedad de Chagas/transmisión , Ensayo de Inmunoadsorción Enzimática/estadística & datos numéricos , Factores de RiesgoRESUMEN
Se presentan los resultados de una prueba diagnóstica novedosa, basada en la amplificación de una secuencia nucleótida conocida de un ADN blanco. La reacción de amplificación génica (PCR), descrita en este trabajo, permite una amplificación * 10 elevado a 6 -veces un segmento de 294 bp de un gene que codifica a un antígeno de 65 KDa- correspondiente a una proteína de respuesta al estrés térmico, presente en Mycobacterium tuberculosis y Mycobacterium bovis BCG. Esta reacción requiere grandes cantidades de ADN blanco, pero con la técnica utilizada fue posible detectar amplificación comenzando con 10 elevado a -12 g de ADN; cantidad que corresponde a 10-100 micobacterias/equivalentes ADN. Este nivel detectado representa un mejoramiento respecto a la tinción Ziehl-Neelsen, el cual necesita sobre 10.000 micobacterias para ser leído como positivo. La concentración de Mg elevado a +2 es crítica en la reacción, la cual corresponde a 3-5 mM. El análisis de la secuencia muestra que la amplificación de ésta es comparable a la secuencia localizada entre las posiciones 781-1075 del gene 65 KDa. Se realizaron pruebas específicas que demostraron que la señal de amplificación es posible de encontrar con ADN obtenido de M. tuberculosis y M. bovis BCG. Con otras micobacterias la señal de amplificación es muy baja y equivalente a aquéllas encontradas con ADN micobacterial no relacionado. La reacción fue probada con cuatro muestras clínicas, las cuales fueron positivas para la presentación de micobacterias; en tres de ellas se encontró el segmento amplificado 294 bp. Continúan desarrollándose ensayos de sensibilidad y especificidad en muestras clínicas