Application of extended single-reaction multiplex polymerase chain reaction for toxin typing of Staphylococcus aureus isolates in South Korea

• Published in: International journal of food microbiology Vol. 97; no. 2; pp. 137 - 145
• Main Authors: Kwon, N.H., Kim, S.H., Park, K.T., Bae, W.K., Kim, J.Y., Lim, J.Y., Ahn, J.S., Lyoo, K.S., Kim, J.M., Jung, W.K., Noh, K.M., Bohach, G.A., Park, Y.H.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15-12-2004; Elsevier
• Subjects: Abattoirs; AP-PCR; Bacterial Typing Techniques; Biological and medical sciences; DNA Fingerprinting; DNA Primers; DNA, Bacterial - genetics; DNA, Bacterial - isolation & purification; Enterotoxins - genetics; Enterotoxins - isolation & purification; Food industries; Food Microbiology; Fundamental and applied biological sciences. Psychology; Multiplex PCR; Polymerase Chain Reaction - methods; Staphylococcal enterotoxin; Staphylococcal Food Poisoning - prevention & control; Staphylococcus aureus; Staphylococcus aureus - classification; Staphylococcus aureus - isolation & purification; Staphylococcus aureus - metabolism; Asia; Korea; Korea, Rep; Multiplex PCR; AP-PCR; Staphylococcal enterotoxin; Staphylococcus aureus; Typing; Multiplex polymerase chain reaction; Random amplified polymorphic DNA marker; Polymerase chain reaction; Toxin; Enterotoxin; Bacteria; Micrococcales; Isolate; Micrococcaceae; Molecular biology; Application
• ISSN: 0168-1605; 1879-3460
• DOI: 10.1016/j.ijfoodmicro.2004.04.014

The extended single-reaction multiplex PCR (esr-mPCR) developed in this study to detect staphylococcal enterotoxins (SEs), including SEA, SEB, SEC, SED, SEE, SEH, SEI, and SEJ, requires fewer sets of primers than other conventional multiplex PCRs and can be used to detect newly identified staphylococcal enterotoxins SEs more readily. Esr-mPCR analysis of 141 isolates of
Staphylococcus aureus obtained from abattoir and livestock product samples revealed that 27 of the
S. aureus isolates were toxigenic, and two were 2 multitoxigenic isolates. The most prevalent SE type was SEI followed by SEA and SEH. In addition, we investigated the clonal relatedness of toxigenic
S. aureus isolates by arbitrarily primed PCR (AP-PCR). AP-PCR analysis of toxigenic
S. aureus isolates revealed that the discriminatory power of AP-PCR was 9 (
D=0.81), 8 (
D=0.77), and 10 types (
D=0.83) with primers AP1, ERIC2, and AP7, respectively. The combination of three each AP-PCR result could rearrange toxigenic
S. aureus isolates into 10 types and five subtypes, with the
D-value of 0.92. Interestingly, our data showed that toxigenic
S. aureus isolates from different sources had different fingerprinting patterns although some of them carried the same types of SE genes. These data suggest that combinations of esr-mPCR and AP-PCR can provide a powerful approach for epidemiological investigation of toxigenic
S. aureus isolates.