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Colorimetric DNAzyme Biosensor for Convenience Detection of Enterotoxin B Harboring Staphylococcus aureus from Food Samples
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2018-02-05 00:00:00 , DOI: 10.1021/acs.jafc.7b04820 Bhairab Mondal 1 , Bhavanashri N 1 , Shylaja Ramlal 1 , Joseph Kingston 1
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2018-02-05 00:00:00 , DOI: 10.1021/acs.jafc.7b04820 Bhairab Mondal 1 , Bhavanashri N 1 , Shylaja Ramlal 1 , Joseph Kingston 1
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In the present study, a colorimetric DNAzymes biosensor strategy was devised in combination with immunomagnetic separation for rapid and easy detection of enterotoxin B harboring Staphylococcus aureus from food and clinical samples. The method employs immunocapture of S. aureus and amplification of seb gene by DNAzyme complementary sequence integrated forward primer and with specific reverse primer. The DNAzyme sequence integrated dsDNA PCR products when treated with hemin and TMB (3,3′,5,5′-tetramethylbenzidine) in the presence of H2O2 produce colorimetric signal. A linear relationship of optical signal with the initial template of seb was obtained which could be monitored by visually or spectrophotrometrically for qualitative and quantitative detection. The limit of detection for the assay was approximately 102 CFU/mL of seb gene harboring target. This method is convenient compared to gel based and ELISA systems. Further, spiking studies and analysis on natural samples emphasized the robustness and applicability of developed method. Altogether, the established assay could be a reliable alternative, low-cost, viable detection tool for the routine investigation of seb from food and clinical sources.
中文翻译：
比色法DNAzyme生物传感器，用于方便地检测食物样品中含有金黄色葡萄球菌的肠毒素B
在本研究中，比色DNA酶生物传感器策略被设计与免疫磁分离相结合，用于从食品和临床样品中快速简便地检测出含有金黄色葡萄球菌的肠毒素B。该方法利用金黄色葡萄球菌的免疫捕获，并通过整合有正向引物和特定反向引物的DNAzyme互补序列扩增seb基因。当在H 2 O 2存在下用血红素和TMB（3,3'，5,5'-四甲基联苯胺）处理时，DNAzyme序列整合的dsDNA PCR产物产生比色信号。光信号与seb初始模板的线性关系获得了可以通过视觉或分光光度法进行监测以进行定性和定量检测的产物。该测定的检测极限是约10 2 CFU / mL的seb基因带有靶标。与基于凝胶和ELISA的系统相比，此方法很方便。此外，对天然样品的研究和分析突显了所开发方法的稳健性和适用性。总体而言，已建立的检测方法可以作为一种可靠的替代，低成本，可行的检测工具，用于食品和临床来源的皮脂的常规调查。
更新日期：2018-02-05
中文翻译：
比色法DNAzyme生物传感器，用于方便地检测食物样品中含有金黄色葡萄球菌的肠毒素B
在本研究中，比色DNA酶生物传感器策略被设计与免疫磁分离相结合，用于从食品和临床样品中快速简便地检测出含有金黄色葡萄球菌的肠毒素B。该方法利用金黄色葡萄球菌的免疫捕获，并通过整合有正向引物和特定反向引物的DNAzyme互补序列扩增seb基因。当在H 2 O 2存在下用血红素和TMB（3,3'，5,5'-四甲基联苯胺）处理时，DNAzyme序列整合的dsDNA PCR产物产生比色信号。光信号与seb初始模板的线性关系获得了可以通过视觉或分光光度法进行监测以进行定性和定量检测的产物。该测定的检测极限是约10 2 CFU / mL的seb基因带有靶标。与基于凝胶和ELISA的系统相比，此方法很方便。此外，对天然样品的研究和分析突显了所开发方法的稳健性和适用性。总体而言，已建立的检测方法可以作为一种可靠的替代，低成本，可行的检测工具，用于食品和临床来源的皮脂的常规调查。
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