Abstract
PurposeA real-time immunocapture PCR (RT-IPCR) has been fabricated for the detection of Staphylococcus aureus enterotoxin B (SEB) from food and environmental samples.MethodsConsidering the fact, anti-SEB immunoglobulin G (IgG) has affinity towards protein A, produced by nearly all S. aureus, and generates false-positive read out in all immuno-based assay. We have employed avian anti-SEB antibody (SEB-IgY) as capture probe, since IgY interact less efficiently to protein A and biotinylated SEB-specific monoclonal antibody (SEB -MAb) conjugated with reporter DNA as revealing probe for real-time PCR amplification and signal generation. Sensitivity and selectivity of the assay were evaluated employing closely related enterotoxins and other toxins.ResultsThe RT-IPCR is highly specific and sensitive (100 fg/mL). The practical applicability of the assay was tested using spiked food sample as well as naturally contaminated food samples. The sensitivity and specificity of RT-IPCR were not compromised by the foods tested and was able to detect SEB conveniently. Further, the assay was validated comparing with the in-house developed PCR, and plausible result was obtained.ConclusionThe developed assay can be utilized as a low-cost detection system of SEB in routine food testing laboratories.
Highlights
Superantigenic staphylococcal enterotoxins (SEs) (SEA, Staphylococcus aureus enterotoxin B (SEB), SEC, and remaining until SEV) and toxic shock syndrome toxin (TSST) are considered as the major cause of food-borne illness worldwide (Argudín et al 2010; Ortega et al 2010, and Fratamico et al 2005)
Almost all S. aureus strains produce a number of enterotoxins, but this does not conclude their toxin production capability in food as their toxin-producing ability varies based on the food environment
The Fc domain of immunoglobulin G (IgG) has an affinity to staphylococcal protein A (SpA) and produce falsepositive results in all immunoassays
Summary
Considering the fact, anti-SEB immunoglobulin G (IgG) has affinity towards protein A, produced by most S. aureus, and generates false-positive read out in all immuno-based assay. We have employed avian anti-SEB antibody (SEB-IgY) as capture probe, since IgY interact less efficiently to protein A and biotinylated SEB-specific monoclonal antibody (SEB -MAb) conjugated with reporter DNA as revealing probe for real-time PCR amplification and signal generation. Sensitivity and selectivity of the assay were evaluated employing closely related enterotoxins and other toxins
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
