RESUMEN
The aim of this study was to follow contamination of ready-to-eat food with Listeria monocytogenes by using the Step One real time polymerase chain reaction (PCR). We used the PrepSEQ Rapid Spin Sample Preparation Kit for isolation of DNA and MicroSEQ® Listeria monocytogenes Detection Kit for the real-time PCR performance. In 30 samples of ready-to-eat milk and meat products without incubation we detected strains of Listeria monocytogenes in five samples (swabs). Internal positive control (IPC) was positive in all samples. Our results indicated that the real-time PCR assay developed in this study could sensitively detect Listeria monocytogenes in ready-to-eat food without incubation.
Asunto(s)
Productos Lácteos/microbiología , Microbiología de Alimentos/métodos , Listeria monocytogenes/aislamiento & purificación , Carne/microbiología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Animales , ADN Bacteriano/análisis , ADN Bacteriano/aislamiento & purificación , Microbiología de Alimentos/instrumentación , Juego de Reactivos para Diagnóstico , Reacción en Cadena en Tiempo Real de la Polimerasa/instrumentación , Salmón/microbiología , Alimentos Marinos/microbiología , Sensibilidad y Especificidad , Ovinos/microbiologíaRESUMEN
Chicks of a conventional poultry flock, Shaver Starcross 288 hybrid, were vaccinated with infectious bronchitis (IB) virus H 120 at the age of 21 days. Three weeks later, the chicks were divided into three groups and separate groups were infected with infectious bronchitis viruses M 41 and D 274 or revaccinated with virus H 120. The content of specific antibodies to antigens prepared from homologous and heterologous viruses of infectious bronchitis used for chick vaccination and infection was investigated at regular intervals in the separate groups of chicks by means of an ELISA technique and haemagglutination-inhibition test (HIT). Serotype specificity of haemagglutination-inhibition test was documented by the results; the specificity was obvious mainly after the first vaccination and two weeks after infection, or after chick revaccination (Fig. 1). The dynamics of postinfective or postvaccinal antibodies, recorded by the ELISA technique, had analogical patterns in the separate groups of chicks, and there were no larger differences in the values determined on the basis of different antigens during the investigation (Fig. 2). A total of 52 group samples of fowl serum was examined by the ELISA technique and agar-gel precipitin test (AGPT) in another part of this study. Ten serums of identical origin represented the separate groups. The result of this examination was evaluated from the percentage of samples with precipitin activity in the group, or from the average value of ELISA. Mutual comparison of the mentioned values indicated that the precipitin activity was limited by the positivity degree of ELISA reaction (Fig. 3).(ABSTRACT TRUNCATED AT 250 WORDS)