1. Immunoquantitative Real-Time PCR for Detection and Quantification of Staphylococcus aureus Enterotoxin B in Foods.
- Author
-
Rajkovic, Andreja, El Moualij, Benaissa, Uyttendaele, Mieke, Brolet, Philippe, Zorzi, Willy, Heinen, Ernst, Foubert, Ellen, and Debevere, Johan
- Subjects
- *
STAPHYLOCOCCUS aureus , *ENTEROTOXINS , *STAPHYLOCOCCUS aureus infections , *ENZYME-linked immunosorbent assay , *BACTERIAL toxins , *IMMUNOGLOBULINS , *DNA probes , *FOOD , *IMMUNOENZYME technique - Abstract
A real-time immunoquantitative PCR (iqPCR) method for detection of Staphylococcus aureus enterotoxin B (SEB) was developed and evaluated using both pure cultures and foods. The assay consisted of immunocapture of SEB and real-time PCR amplification of the DNA probe linked to the detection antibody. iqPCR was compared to an in-house enzyme-linked immunosorbent assay (ELISA) using the same couple of capture-detection antibodies and to commercial kits for detection of S. aureus enterotoxins (SE). The iqPCR was approximately 1,000 times more sensitive (<10 pg ml-1) than the in-house ELISA and had a dynamic range of approximately 10 pg ml-1 to approximately 30,000 pg ml-1. iqPCR was not inhibited by any of the foods tested and was able to detect SEB present in these foods. No cross-reactivity with SE other than SEB was observed. Application of iqPCR for detection of SEE in cultures of S. aureus revealed the onset of SEB production after 4 h of incubation at 22, 37, and 42°C, which was in the first half of the exponential growth phase. The total amounts of SEB produced by the two strains tested were larger at 42°C than at 37°C and were strain dependent. [ABSTRACT FROM AUTHOR]
- Published
- 2006
- Full Text
- View/download PDF