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A G-quadruplex DNAzyme-based LAMP biosensing platform for a novel colorimetric detection of Listeria monocytogenes
Analytical Methods ( IF 3.1 ) Pub Date : 2018-01-15 00:00:00 , DOI: 10.1039/c7ay02908j
Zhanmin Liu 1, 2, 3, 4 , Chenhui Yao 1, 2, 3, 4 , Yanming Wang 1, 2, 3, 4 , Cuiyun Yang 4, 5, 6
Affiliation  

Listeria monocytogenes is one of the most common pathogenic bacteria. After infection, the main symptoms of listeriosis are sepsis, meningitis and mononucleosis. A G-quadruplex DNAzyme-based LAMP biosensing platform was developed for the colorimetric detection of L. monocytogenes. Four primers were designed to hybridize against six distinct sequences in the target DNA making the highly specific amplification in the step of LAMP. The G-quadruplex sequence amplified simultaneously in LAMP could form DNAzyme with the addition of hemin. The catalytic horseradish peroxidase-mimicking DNAzymes allowed the colorimetric responses of the target DNA from L. monocytogenes. The response surface methodology was used to screen and optimize the crucial factors affecting the response. This assay could detect L. monocytogenes specifically with as low as 47.5 CFU per reaction by the naked eye. Furthermore, this colorimetric detection assay was successfully applied to detect L. monocytogenes in spiked pork samples. Therefore, the rapid, specific and visual detection of L. monocytogenes has demonstrated potentially broad applicability in food safety.

中文翻译:

一个基于G-四链体DNA酶的LAMP生物传感平台,用于单核细胞增生李斯特菌的比色检测

单核细胞增生李斯特菌是最常见的致病菌之一。感染后,李斯特菌病的主要症状是败血症,脑膜炎和单核细胞增多症。开发了一个基于G-四链体DNA酶的LAMP生物传感平台,用于比色法检测单核细胞增生李斯特菌。设计了四种引物以与目标DNA中的六个不同序列杂交,从而在LAMP步骤中进行了高度特异性的扩增。在LAMP中同时扩增的G-四链体序列可在添加血红素的情况下形成DNA核酶。催化辣根过氧化物酶模拟DNA酶可以使单核细胞增生李斯特氏菌靶DNA的比色反应。响应面方法用于筛选和优化影响响应的关键因素。该测定法用肉眼每次反应可低至47.5 CFU地特异性检测单核细胞增生李斯特菌。此外,该比色检测法已成功应用于检测加标猪肉样品中的单核细胞增生李斯特氏菌。因此,对单核细胞增生李斯特氏菌的快速,特异性和视觉检测已证明在食品安全中具有潜在的广泛适用性。
更新日期:2018-01-15
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