Detecção simultânea de Salmonella spp. e Salmonella enteritidis em carcaças de frango por reação em cadeia da polimerase

AUTOR(ES)

Daniel Santos Pinto Silva

DATA DE PUBLICAÇÃO

2008

RESUMO

An increase in human salmonellosis caused by serovar Enteritidis related to consumption of poultry products was observed worldwide in the last years. Safety measures have been adopted to control this serovar. The implementation of Hazard Analysis and Critical Control Points (HACCP) System was not able to eliminate the problem although it has considerably reduced the number of infected chickens. Salmonellosis is the most frequent bacterial foodborne disease in Parana State, Brazil and serovar Enteritidis caused approximately 80% of outbreaks occurred between 1999 and 2006. S. Enteritidis causes subclinical infection in chickens. Thus, the constant checking of Salmonella contamination in the food chain is essential to control human disease. Salmonella detection in food, water or environmental samples by traditional methods is labor-intensive and time-consuming. Molecular methods, such as polymerase chain reaction (PCR), are rapid, specific and sensitive and several methods have been developed to detect and identify Salmonella in foods. The objective of this study was to develop a multiplex PCR for detection of Salmonella spp. and Salmonella Enteritidis in chicken carcasses. A PCR was developed using Salmonella genus-specific primers INVA and serovar Enteritidis-specific primers IE1, which produce specific DNA fragments of 796 and 316 base-pairs, respectively. The standardized PCR was tested with different isolates of Salmonella and other bacterial species and the specificity was 100%. Salmonella was detected from chicken skin artificially contaminated with approximately 1 colonyforming unit (CFU) of Salmonella per 10 g, after 24 h of non-selective enrichment. Salmonella spp. was detected in nine (13%) of 66 analyzed chicken carcasses purchased from local retail market. The presence of Salmonella was confirmed by traditional culture method and serovars Schwarzengrund and Montevideo were identified in six and three carcass samples, respectively. Specific DNA fragments of the PCR confirmed the presence of genus Salmonella, but not serovar Enteritidis. The standardized multiplex PCR was found to be sensitive and specific on the detection of genus Salmonella and simultaneous identification of serovar Enteritidis in chicken meat after 24 hours of analysis.

ASSUNTO(S)

salmonela enteritidis salmonela spp alimentos - microbiologia ave - carcaça - doenças food - microbiology poultry - carcasses - diseases

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