Introduccion: las enfermedades transmitidas por alimentos constituyen un problema mundial de salud publica y generan negativo impacto socioeconomico. Objetivo: identificar el agente causal de un brote de intoxicacion alimentaria en un comedor escolar. Material y metodos: estudio realizado en afectados por una intoxicacion alimentaria en el municipio de Santo Tomas, departamento del Atlantico, Colombia, en el primer semestre de 2008. Se realizo identificacion fenotipica de 24 cepas de Staphylococcus aureus , aisladas de vomito, materia fecal, jugo gastrico, secrecion de lesion en dedo, secrecion nasofaringea y faringea en escolares y manipuladores de alimento. De estas cepas, 15 se analizaron por medio de la tecnica de PCR-RAPD (amplificacion aleatoria de ADN polimorfico (random amplification of polimorphic DNA-RAPD). Los productos de PCR se visualizaron en geles de agarosa, se analizaron por medio de un dendograma y se calculo el coeficiente de similitud de Jaccard. Resultados: en 16 muestras de materia fecal se identifico Staphylococcus aureus coagulasa positiva y en dos Salmonella spp. En vomito 6 fueron positivas para S. aureus coagulasa positiva y una para Salmonella spp. Tambien en una muestra de secrecion de una y en una toma nasofaringea se aislo S. aureus coagulasa positivo. Conclusion: se identificaron cepas de S. aureus con amplia diversidad molecular, y constituye el posible agente causal de un brote por intoxicacion alimentaria en un comedor escolar. Rev.cienc.biomed. 2013;4(2):233-241 PALABRAS CLAVE Staphylococcus aureus; Brotes de enfermedades; Intoxicacion alimentaria; Salmonella, Tecnica del ADN polimorfo amplificado aleatorio. SUMMARY Introduction: foodborne diseases constitute a worldwide problem of public health and generate a negative socio-economic impact. Objective: to identify the causal agent of an outbreak of food poisoning in a school cafeteria. Methods: Study carried out in people affected by a food poisoning in the municipality of Santo Tomas, Atlantico, Colombia, in the first semester of 2008. Phenotypic identification of 24 families of Staphylococcus aureus isolated from vomiting, fecal material, gastric juices, secretion of injury in finger and nasopharyngeal and pharyngeal secretion in schoolchildren and food handlers was carried out. Of these families, 15 were analyzed by means of PCR-RAPD (Random Amplified Polymorphic DNA) technique. The products of PCR were visualized in agarose gels and analyzed by means of a dendogram. The similarity coefficient of Jaccard was calculated. Results: Coagulase-positive Staphylococcus aureus was identified in 16 samples of fecal material and Salmonella spp was recognized in two samples. In vomiting, 6 samples were positive for Coagulase-positive S. aureus and one sample for Salmonella spp. Also, Coagulase-positive S. aureus was isolated in a sample of secretion of nail and in nasopharyngeal secretion. Conclusion: Families of S. aureus with wide molecular diversity were identified. It constitutes the possible causal agent of an outbreak by food poisoing in a school cafeteria. Rev.cienc.biomed. 2013;4(2):233-241 KEYWORDS Staphylococcus aureus; Outbreaks; Food poisoning; Salmonella; Random Amplified Polymorphic DNA technique.