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Electrochemical immunosensor for the detection of staphylococcal enterotoxin B using screen-printed electrodes
Indian Journal of Chemistry, Section A ( IF 0.412 ) Pub Date : 2020-02-14
Arun Sharma, Vepa Kameswara Rao, Dev Vrat Kamboj

Staphylococcal enterotoxin B (SEB) is responsible for large number of food poisoning cases throughout the world. SEB is an exotoxin and it is one of the several harmful substances produced by the bacterium Staphylococcus aureus. Therefore, it is required to develop methods for the sensitive, reliable, reproducible, cheaper, easy to use and rapid detection of SEB. An electrochemical immunosensor has been studied for the fast detection of SEB using disposable screen-printed electrodes. Ascorbic acid-2-phosphate (AA-2P) is used as a new substrate for the voltammetric detection of SEB. In the alkaline buffer solution the alkaline phosphate (ALP) enzymatic hydrolysis product of AA-2P is ascorbic acid (AA). Ascorbic acid is an electroactive substance and gives differential pulse voltammetric oxidative response at +380 mV (versus Ag/AgCl). The potential +0.39 V corresponds to the oxidation of AA. Indirect sandwiched enzyme linked immunosorbent assay (ELISA) has been used for the detection of SEB. In this method, anti-rabbit IgG for SEB (capturing antibody) is first immobilized on the surface of SPE followed by reacting with SEB (antigen) to form antigen-antibody complex. After that, anti-mice IgG for SEB (secondary antibody) are added, followed by ALP-conjugated anti-mice IgG (revealing antibody). The optimal conditions for ALP enzymatic reaction and the volumetric detection have been optimized. It is found that the response of voltammetric immunosensor is proportional to the SEB concentration in the range 0.1–100 ng/mL and the detection limit was found to be 0.1 ng/mL.

中文翻译:

使用丝网印刷电极检测葡萄球菌肠毒素B的电化学免疫传感器

葡萄球菌肠毒素B(SEB)导致全世界大量食物中毒案件。SEB是一种外毒素,它是金黄色葡萄球菌细菌产生的几种有害物质之一。。因此,需要开发灵敏,可靠,可再现,便宜,易于使用和快速检测SEB的方法。已经研究了一种电化学免疫传感器,用于使用一次性丝网印刷电极快速检测SEB。抗坏血酸-2-磷酸酯(AA-2P)被用作SEB伏安检测的新底物。在碱性缓冲溶液中,AA-2P的碱性磷酸盐(ALP)酶水解产物是抗坏血酸(AA)。抗坏血酸是一种电活性物质,在+380 mV(相对于Ag / AgCl)下产生微分脉冲伏安氧化反应。+0.39 V电位对应于AA的氧化。间接夹心酶联免疫吸附测定(ELISA)已用于检测SEB。用这种方法 首先将SEB(捕获抗体)的抗兔IgG固定在SPE的表面,然后与SEB(抗原)反应形成抗原-抗体复合物。之后,添加用于SEB的抗小鼠IgG(二抗),然后添加ALP缀合的抗小鼠IgG(揭示抗体)。优化了ALP酶促反应和体积检测的最佳条件。发现伏安免疫传感器的响应与SEB浓度成正比,范围为0.1–100 ng / mL,检测极限为0.1 ng / mL。优化了ALP酶促反应和体积检测的最佳条件。发现伏安免疫传感器的响应与SEB浓度成正比,范围为0.1–100 ng / mL,发现极限为0.1 ng / mL。优化了ALP酶促反应和体积检测的最佳条件。发现伏安免疫传感器的响应与SEB浓度成正比,范围为0.1–100 ng / mL,发现极限为0.1 ng / mL。
更新日期:2020-02-14
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