环介导等温扩增法快速检测耐甲氧西林金黄色葡萄球菌-食品科学.pdfVIP

※分析检测 食品科学 2011, Vol. 32, No. 16 213 朱 丽，霍贵成 * (东北农业大学 乳品科学教育部重点实验室，黑龙江 哈尔滨 150030) 建立一种应用环介导等温扩增(loop-mediated isothermal amplification ，LAMP)方法快速检测耐甲氧西林金黄 色葡萄球菌(methicillin-resistan Staphylococcus aureus ，MRSA) 中mecA 和spa 基因的检测方法。以金葡菌和其他相 关菌种为试验对象，分别用聚合酶链式反应(ploymerase chain reaction，PCR)和 LAMP 方法检测 mecA 和spa 基因。 结果表明：LAMP 法在 64 ℃等温条件下可在60min 内成功扩增基因，且与传统PCR 方法结果相同；通过琼脂糖凝 2 胶电泳可知，LAMP 对 mecA 和spa 基因的检测限分别为每菌管 10 个和 10 个细胞；肉眼可检测到的 mecA 和spa 基 3 个和 10 个细胞；然后用 LAMP 法检测人工污染原料乳样本中 MRSA ，用LAMP 法检 因的检测限分别为每菌管 10 测原料乳中的 mecA 和spa 基因与 PCR 方法显示出相同的结果。LAMP 方法可快速检测 mecA 和spa 基因，此方法可 应用于原料乳中 MRSA 的检测。 环介导等温扩增(LAMP) ；mecA 基因；耐甲氧西林金黄色葡萄球菌(MRSA) ；肉眼检测；spa 基因 Rapid Detection of Methicillin-resistant Staphylococcus aureus by Loop-mediated Isothermal Amplification ZHU Li ，HUO Gui-cheng* (Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University, Harbin 150030, China) Abstract：A rapid assay was developed for the detection of mecA and spa gene present in methicillin-resistant Staphylococcus aureus (MRSA) using loop-mediated isothermal amplification (LAMP). Staphylococcus aureus and other relevant bacteria were detected for their mecA and spa genes by both polymerase chain reaction (PCR) and LAMP methods. Gene amplification was successfully achieved by LAMP within 60 min when the temperature was held at 64 ℃, and the results were consistent results with those of the conventional PCR methods. The limits of detection of the LAMP method for mecA and spa genes 2