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Multiplex PCR ( mPCR ) for the Detection of S almonella spp. and the Differentiation of the T yphimurium and E nteritidis Serovars in Chicken Meat
Author(s) -
Saeki Erika Kushikawa,
Alves Juliane,
Bonfante Raissa Curti,
Hirooka Elisa Yoko,
Oliveira Tereza Cristina Rocha Moreira
Publication year - 2013
Publication title -
journal of food safety
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.427
H-Index - 43
eISSN - 1745-4565
pISSN - 0149-6085
DOI - 10.1111/jfs.12019
Subject(s) - serotype , multiplex polymerase chain reaction , microbiology and biotechnology , salmonella enterica , polymerase chain reaction , biology , real time polymerase chain reaction , salmonella , gene , food science , bacteria , genetics
almonella enterica subspecies enterica serovars Enteritidis and Typhimurium are important causes of foodborne illness. Methods for simultaneous detection of these serovars may contribute for the adoption of measures to prevent these diseases. The polymerase chain reaction to detect individually or simultaneously the serovars Enteritidis and Typhimurium in foods have been standardized; however, the majority of assays employ the fli C gene as a target for detection of serovar Typhimurium. The detection of these sorovars in a few hours allows the food supply chain to take appropriate measures to prevent the distribution of contaminated food. The aim of this study was to develop a new multiplex PCR ( mPCR ) for the simultaneous detection and differentiation of S almonella spp., S . Enteritidis and S . Typhimurium in chicken meat. The mPCR assays showed high specificity and differentiated S . Typhimurium from 22 S almonella serovars tested, including S . K entucky, showing that it's an alternative to reduce the time required to obtain presumptive positive results. Practical Applications The fli C gene used as target for the detection of serovar T yphimurium is questionable as it has also been described for S . K entucky. The amplification of STM 4492 gene in this study, instead of fli C gene, exhibited high specificity to detect S . Typhimurium. The developed mPCR is an efficient means for the simultaneous detection and differentiation of S almonella spp., S . Enteritidis and S . Typhimurium in chicken meat after 24 h of enrichment. The mPCR has the potential to be used in routine diagnostic laboratories to obtain presumptive positive results and for identification of E nteritidis and T yphimurium strains isolated by the conventional method.