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Colorimetric aptasensor for detecting Salmonella spp., Listeria monocytogenes, and Escherichia coli in meat samples
Journal of Near Infrared Spectroscopy ( IF 1.6 ) Pub Date : 2020-01-16 , DOI: 10.1177/1082013219899593
Sudarat Ledlod 1, 2, 3 , Supatra Areekit 4, 5 , Somchai Santiwatanakul 5, 6 , Kosum Chansiri 1, 5
Affiliation  

In this study, we successfully developed a simple and rapid method for simultaneous detection of Salmonella spp., Listeria monocytogenes, and Escherichia coli using gold nanoparticles and the aptamer aptasensor. We screened 25 specific DNA aptamer candidates against these pathogens using whole-cell Systematic Evolution of Ligands by EXponential enrichment. Among them, Ap6 was selected due to its low energy minimization values of −12.25 and −27.67 kcal/mol derived from MFold and RNAFold analysis, respectively. The assay presented in this study allowed the visual colorimetric detection of labeled colloidal gold nanoparticles as well as determination of UV absorbance at 625 and 525 nm under optimized conditions. The detection limit of this aptasensor was as less as 105 CFU/ml. A random investigation of 50 meat samples, including ham and chicken sausages, collected from the local market revealed 96% accuracy, 96% specificity, and 100% sensitivity of the assay. The colorimetric aptasensor can accomplish one-step detection without pre-culture, DNA extraction, and amplification. Hence, it is an easy, rapid, specific, and qualitative assay that can be used as a point-of-care testing to directly detect multiplex foodborne pathogens.



中文翻译:

比色适体传感器,用于检测肉样品中的沙门氏菌单核细胞增生李斯特菌大肠杆菌

在这项研究中,我们成功开发了一种使用金纳米颗粒和适体适体传感器同时检测沙门氏菌单核细胞增生李斯特菌大肠杆菌的简单快速方法。我们使用的全细胞筛选针对这些病原体25特定DNA适配子候选Ş ystematic Ë的旋涡大号由igands EX潜在浓缩。其中,选择Ap6的原因是其分别来自MFold和RNAFold分析的低能量最小化值-12.25和-27.67 kcal / mol。在这项研究中提出的测定方法允许视觉上比色法检测标记的胶体金纳米颗粒,以及在最佳条件下测定625和525 nm处的紫外线吸收率。该适体传感器的检出限小于10 5CFU /毫升。从当地市场上对包括火腿和鸡肉香肠在内的50种肉类样品进行的随机调查显示,该检测方法的准确度为96%,特异性为96%,灵敏度为100%。比色适体传感器无需预先培养,DNA提取和扩增即可完成一步检测。因此,它是一种简便,快速,特异性和定性的测定方法,可用作直接检测多种食源性病原体的即时检验。

更新日期:2020-04-10
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