A real-time immunocapture PCR (RT-IPCR) has been fabricated for the detection of Staphylococcus aureus enterotoxin B (SEB) from food and environmental samples. Considering 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. The 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. The developed assay can be utilized as a low-cost detection system of SEB in routine food testing laboratories.

中文翻译：

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实时免疫捕获PCR（RT-IPCR），无蛋白A干扰，可从食品和环境样品中方便地检测葡萄球菌肠毒素B

实时免疫捕获PCR（RT-IPCR）已被制成用于从食物和环境样品中检测金黄色葡萄球菌肠毒素B（SEB）。考虑到这一事实，抗SEB免疫球蛋白G（IgG）对几乎由所有金黄色葡萄球菌产生的蛋白A具有亲和力，并在所有基于免疫的测定中产生假阳性读数。我们已使用禽类抗SEB抗体（SEB-IgY）作为捕获探针，因为IgY与蛋白质A和与报道基因DNA偶联的生物素化SEB特异性单克隆抗体（SEB -MAb）的相互作用效率较低，是实时PCR扩增的揭示探针和信号生成。使用紧密相关的肠毒素和其他毒素评估了测定的灵敏度和选择性。RT-IPCR具有高度特异性和敏感性（100 fg / mL）。使用加标食品样本和自然污染食品样本测试了该测定方法的实际适用性。RT-IPCR的敏感性和特异性不受测试食物的影响，能够方便地检测SEB。此外，与内部开发的PCR相比，该测定法得到了验证，并获得了合理的结果。所开发的检测方法可以在常规食品检测实验室中用作SEB的低成本检测系统。