Development of a multiplex PCR assay for the identification of Staphylococcus aureus enterotoxigenic strains isolated from milk and dairy products.
A multiplex PCR for the simultaneous detection of Staphylococcus aureus 23S rRNA, the coagulase and thermonuclease genes as well as the enterotoxin genes sea, sec, sed, seg, seh, sei, sej, sel was developed. The method was used to determine the presence of enterotoxigenic types for 93 S. aureus strains isolated from milk and dairy products. The data obtained by mPCR resulted comparable to those obtained by immunoassay methods. In addition, the mPCR assays also amplified some se genes, whose toxins are undetectable by immunoassay. Multiplex amplification can be obtained starting from 1 pg of DNA, showing the excellent specificity and high sensitivity of the assay.
Cremonesi
Paola
P
Department of Biomedical Sciences and Technologies, University of Milan, Via Fratelli Cervi 93, Segrate, Milan, Italy.
Luzzana
Massimo
M
Brasca
Milena
M
Morandi
Stefano
S
Lodi
Roberta
R
Vimercati
Chiara
C
Agnellini
Dario
D
Caramenti
Giancarlo
G
Moroni
Paolo
P
Castiglioni
Bianca
B
D000818:Animals; D003611:Dairy Products / Q000382:microbiology*; D004768:Enterotoxins; D005506:Food Contamination; D006801:Humans; D008892:Milk / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012338:RNA, Ribosomal, 23S / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000145:classification / Q000235:genetics*
D004768:Enterotoxins; D012338:RNA, Ribosomal, 23S