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Designing of polymerase chain reaction primers for the detection of Salmonella enteritidis in foods and faecal samples
Author(s) -
Wang S.J.,
Yeh D.B.
Publication year - 2002
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.1472-765x.2002.01114.x
Subject(s) - salmonella enteritidis , polymerase chain reaction , biology , salmonella , primer (cosmetics) , microbiology and biotechnology , real time polymerase chain reaction , gene , bacteria , genetics , chemistry , organic chemistry
Aims: In this study, novel insertion element (IE) DNA targeted polymerase chain reaction (PCR) primers were designed and further used for the specific detection of Salmonella enteritidis in foods and faecal samples. Methods and Results: Polymerase chain reaction primers, based upon their IE gene sequence (accession number Z83734), were developed for the detection of Salm. enteritidis . These primers were termed IE1L‐IE1R and IE2L‐IE3R. The cell lysate, rather than the extracted DNA, was used as template and preculturing of bacterial material was carried out prior to the PCR assay. The specificities of these developed primers were to be confirmed further. The PCR procedure developed was used to examine 170 endogenously contaminated samples, including poultry, seafood, meats, faecal specimens and some feed samples. Salmonella enteritidis was detected in 5·29% (nine of 170) of the samples. Conclusions, Significance and Impact of the Study: Two sets of novel PCR primers, based upon their IE gene sequence, have been developed. These primers demonstrated the ability to be used for the specific detection of Salm. enteritidis . When PCR primers IE1L‐IE1R were used for the detection of artificially Salm. enteritidis ‐contaminated food samples, as few as 1 cell ml −1 sample could be detected using this PCR process.