Rapid detection of enterotoxigenic Staphylococcus aureus harbouring genes for four classical enterotoxins, SEA, SEB, SEC and SED, by loop-mediated isothermal amplification assay.
OBJECTIVE
The aim of this study was to develop a loop-mediated isothermal amplification (LAMP) assay targeting the genes for the four classical enterotoxins, SEA, SEB, SEC and SED, in Staphylococcus aureus.


RESULTS
Specific primers were designed which target each specific sequence of the enterotoxin genes. With 30 strains of Staph. aureus, the results of the LAMP assay to each enterotoxin, SEA, SEB, SEC and SED, completely accorded with the results of polymerase chain reaction (PCR) assay. Enterotoxin production, determined by a reverse passive latex agglutination assay, strongly correlated with the presence of the corresponding genes. Amplification was not observed when 14 strains of nonenterotoxigenic Staph. aureus and 20 strains consisting of 19 bacterial species other than Staph. aureus were tested. In addition, the sensitivity of the LAMP assay was generally higher than that of conventional PCR assay and it rapidly detected enterotoxigenic Staph. aureus strains within 60 min.


CONCLUSIONS
The LAMP assay developed in this study is rapid, specific and sensitive for the detection of enterotoxigenic Staph. aureus.


CONCLUSIONS
The method is suitable for clinical diagnosis and food safety applications.
Goto
M
M
Division of Microbiology, National Institute of Health Sciences, Setagaya, Tokyo, Japan.
Hayashidani
H
H
Takatori
K
K
Hara-Kudo
Y
Y
D001427:Bacterial Toxins / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers; D004768:Enterotoxins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000235:genetics* / Q000378:metabolism; D018089:Superantigens / Q000235:genetics* / Q000378:metabolism; D013997:Time Factors
D001427:Bacterial Toxins; D017931:DNA Primers; D004768:Enterotoxins; D018089:Superantigens; C019368:enterotoxin D, Staphylococcal; C026066:enterotoxin A, Staphylococcal; C031818:enterotoxin B, staphylococcal; C034959:enterotoxin C, staphylococcal