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首页> 外文会议>International Conference on Medical Engineering and Bioinformatics >A sensitive and rapid detection of Escherichia coli O157 in raw milk by loop-mediated isothermal amplification combined with 4-(2-pyridylazo) resorcinol monosodium salt
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A sensitive and rapid detection of Escherichia coli O157 in raw milk by loop-mediated isothermal amplification combined with 4-(2-pyridylazo) resorcinol monosodium salt

机译：通过环介导的等温扩增与4-（2-吡啶基氮杂）间苯二酚单钠盐组合的原料乳中大肠杆菌O157的敏感和快速检测

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Loop-mediated isothermal amplification (LAMP) allows for the rapid amplification of nucleic acids under isothermal conditions. With the addition of 4-(2-pyridylazo) resorcinol monosodium salt we have developed an efficient, field-friendly procedure for the detection of Escherichia coli 0157. In this report, LAMP was performed at 65°C for 40 min, and the result was visually assessed via a color change from orange to light yellow produced by the reaction of 4-(2-pyridylazo) resorcinol monosodium salt and Mn~(2+) with pyrophosphate realized during the DNA amplification. The detection limit of E. coli O157 by this assay was equivalent to 3-5 cfu. The total assay time, including 15 min for rapid DNA extraction, was approximately 40 min. Data on naturally contaminated raw milk samples indicated that the method was highly specific and sensitive, giving 100% concordance with the ISO 16654:2001 reference method. The results showed that the assay had the advantages of speed and greater sensitivity and with less dependence on equipment than the standard polymerase chain reaction can be useful in the field as a routine diagnostic tool.
机译：环介导的等温扩增（灯）允许在等温条件下快速扩增核酸。加入4-（2-吡啶胺）间苯二酚单钠盐，我们已经开发了一种有效的，现场友好的方法，用于检测大肠杆菌0157.在本报告中，灯在65℃下进行40分钟，结果通过从橙色的颜色变化视觉评估，通过4-（2-吡啶氧基）间苯二酚盐和Mn〜（2+）与在DNA扩增期间实现的焦磷酸盐产生的反应产生的浅黄色。该测定法通过该测定的检测极限相当于3-5 CFU。用于快速DNA提取的总测定时间，包括15分钟，约为40分钟。天然污染的原牛奶样品的数据表明该方法具有高度特异性和敏感性，与ISO 16654：2001参考方法提供100％的一致性。结果表明，该测定具有速度的优点，敏感性更大，而且对设备的依赖性较少，而不是标准的聚合酶链反应可用于常规诊断工具。

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