Novel multiplex PCR for the detection of the Staphylococcus aureus superantigen and its application to raw meat isolates in Korea.
A multiplex PCR assay that allows for the rapid screening of the 19 genes that encode staphylococcal enterotoxins (SEs) (sea to see, and seg to sei), SE-like (SEl) toxins (sej to ser, and seu), and toxic shock syndrome toxin-1 (TSST-1) (tst) was developed in this study. These toxins are included in the pyrogenic toxin superantigen (PTSAg) family and are responsible for many diseases such as staphylococcal food poisoning (SFP) and TSS. The primers were designed based on dual priming oligonucleotide (DPO) technology to detect all of the 19 SAg genes in three sets of PCR. The developed multiplex PCR was applied to 143 Staphylococcus aureus strains isolated from pork and chicken meat in Korea. Almost 50% of the strains possessed at least one of the 19 SAg genes. The most frequently found genes were seg, sei, sem, and sen (53 isolates, 37%), which were often found simultaneously in the same isolate. In those isolates, the seo (39 isolates, 27%) or seu (6 isolates, 4%) genes were frequently found together and this combination (seg, sei, sem, sen, and seo or seu) was considered to be a part of the enterotoxin gene cluster (egc). The sea gene (10 isolates, 7%) was the gene most frequently detected out of all the classical SE genes (sea to see). Although these classical SEs are considered to be major etiological factors in SFP, newly described SE or SEl genes (seg to ser, and seu) were more frequently detected than the classical SE genes in this study. There was no isolate detected containing the seb, sec, sek, sel, or seq genes. S. aureus possessing mobile genetic elements known to encode these SAg genes, such as egc, were presumed to be widely distributed among pork and chicken meats in Korea. The multiplex PCR developed in this study could be applied to the investigation of SAg genes in S. aureus strains isolated from various sources.
Hwang
Sun Young
SY
Department of Microbiology, College of Veterinary Medicine and BK21 Program for Veterinary Science, Seoul National University, Sillim-dong, Gwanak-gu, Seoul 151-742, Republic of Korea.
Kim
So Hyun
SH
Jang
Eun Joo
EJ
Kwon
Nam Hoon
NH
Park
Young Kyung
YK
Koo
Hye Cheong
HC
Jung
Woo Kyung
WK
Kim
Jun Man
JM
Park
Yong Ho
YH
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D003257:Consumer Product Safety; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D004768:Enterotoxins / Q000235:genetics / Q000302:isolation & purification; D005506:Food Contamination / Q000032:analysis*; D006801:Humans; D007723:Korea; D008460:Meat / Q000382:microbiology*; D005810:Multigene Family; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D013202:Staphylococcal Food Poisoning / Q000382:microbiology*; D013211:Staphylococcus aureus / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D018089:Superantigens / Q000235:genetics* / Q000302:isolation & purification; D013552:Swine
D017931:DNA Primers; D004269:DNA, Bacterial; D004768:Enterotoxins; D018089:Superantigens