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投稿时间：2019-03-19
投稿时间：2019-03-19
中文摘要: 为建立食品中金黄色葡萄球菌、单核增生李斯特氏菌的双重快速检测方法。用缓冲蛋白胨水（buffered peptone water，BPW）对两种目标菌进行增菌培养，采用直接提取法提取样本DNA，比较实时聚合酶链式反应体系（real-time polymerase chain reaction，RT-PCR）中的退火温度，调整高分辨率熔解曲线（high-resolution melting curve，HRM）分析条件，建立同时检测金黄色葡萄球菌、单核增生李斯特氏菌的方法。结果表明，PCR 扩增最佳退火温度为58 ℃，两种目标菌的Tm 值分别为金黄色葡萄球菌77.05 ℃，单核增生李斯特氏菌79.39 ℃；试验灵敏度分别可以达到102、103 CFU/g。
Abstract:Establish a method to rapidly detect Staphylococcus aure and Listeria monocytogenes by highresolution melting curve.Use the buffered peptone water to enrich 2 pathogenic bacteria，and the DNA was extracted by direct extraction method.Compared the annealing temperature of real-time polymerase chain reaction（RT-PCR）and adjusted the condition of high-resolution melting to find the same suitable PCR system to detected the 2 bacteria.The method showed that the suitable annealing temperature was 58 ℃；the Tm values of Staphylococcus aure and Listeria monocytogenes were 77.05 ℃ and 79.39 ℃；the sensibilities were 102 CFU/g and 103 CFU/g.
keywords: real-time polymerase chain reaction（RT-PCR） high-resolution melting curve Staphylococcus aureus Listeria monocytogenes duplex reaction
文章编号：202004034 中图分类号： 文献标志码：
基金项目:
| Author Name | Affiliation |
| YAO Yan-ling | Jiaxing Testing Institute for Food and Drug Control，Jiaxing 314050，Zhejiang，China |
引用文本：
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