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Evaluation of a Multiplex PCR assay for simultaneous identification of Salmonella sp., Salmonella Enteritidis and Salmonella Typhimurium from environmental swabs of poultry houses
Author(s) -
Soumet C.,
Ermel G.,
Rose N.,
Rose V.,
Drouin P.,
Salvat G.,
Colin P.
Publication year - 1999
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1365-2672.1999.00488.x
Subject(s) - salmonella , salmonella enteritidis , salmonella enterica , serotype , biology , multiplex polymerase chain reaction , microbiology and biotechnology , immunomagnetic separation , multiplex , polymerase chain reaction , dna extraction , bacteria , virology , gene , genetics , bioinformatics , biochemistry
A Multiplex PCR‐based assay (m‐PCR) with three sets of primers was developed for the detection of all serotypes of Salmonella enterica and the identification of Salmonella Enteritidis and Salmonella Typhimurium. This method was evaluated against a bacteriological method for the analysis of environmental swabs of poultry houses. Samples were pre‐enriched in phosphate‐buffered peptone water for 24 h and subjected to three different protocols prior to PCR: (i) an immunomagnetic separation using DynabeadsTM anti‐ Salmonella (Dynal); (ii) a DNA extraction procedure using the InstageneTM matrix; (iii) an additional step of culture on an MSRV medium. With protocols 1 and 2, eight positive results were found by PCR and 20 with the bacteriological method. Protocol 3 combining MSRV and PCR gave similar results to those obtained from bacteriological methods and allowed Salmonella detection within 2 days.