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Evaluation of different DNA extraction procedures for the detection of Salmonella from chicken products by polymerase chain reaction
Author(s) -
Soumet C.,
Ermel Gwennola,
Fach P.,
Colin P.
Publication year - 1994
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 - 10.1111/j.1472-765x.1994.tb00458.x
Subject(s) - salmonella , polymerase chain reaction , dna extraction , biology , centrifugation , bacteria , dna , microbiology and biotechnology , enumeration , inoculation , incubation , subspecies , food science , chromatography , chemistry , gene , biochemistry , genetics , paleontology , mathematics , combinatorics , immunology
Polymerase chain reaction (PCR) was used after a short pre‐enrichment culture to detect Salmonella subspecies in chicken fillets. A direct PCR assay performed with chicken meat inoculated with Salmonella Typhimurium produced no PCR products. Six different DNA extraction protocols were tested to recover efficiently Salmonella DNA after a short incubation period. Three of them gave results that were reliable, rapid and sensitive. Successful protocols used Proteinase K and/or a centrifugation step to concentrate the samples. For reliable detection of Salmonella subspecies, a few thousand bacterial cells per ml must be present. To obtain this number of bacterial cells with an inoculation of about one cell in 25 g of ionized food products, it was necessary to incubate samples for at least 10 h before PCR. A larger inoculum of approximately 10 cells in 25 g of ionized food products, required 8 h in culture broth to give positive results by PCR‐based assay.