| 动物饲料中沙门氏菌环介导等温扩增(LAMP)快速诊断方法的建立 |
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| 引用本文： | 孙园园,赵鹏.动物饲料中沙门氏菌环介导等温扩增(LAMP)快速诊断方法的建立[J].中国畜牧兽医,2012,39(1):32-36. |
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| 作者姓名： | 孙园园 赵鹏 |
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| 作者单位： | 镇江出入境检验检疫局,江苏镇江 212008 |
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| 基金项目： | 江苏出入境检验检局科研立项(2010KJ05) |
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| 摘 要： | 根据GenBank公布的沙门氏菌高度保守的fimY蛋白基因序列,利用Primer explorer V4软件设计4条针对沙门氏菌fimY基因的特异性引物,通过对Mg2+浓度、dNTP浓度、反应温度、反应时间、特异性、敏感性、重复性和稳定系及符合率等方面,优化LAMP各种反应条件,建立针对沙门氏菌LAMP快速诊断方法。结果表明,该方法具有较强的特异性;比PCR检测方法敏感性高100倍,对沙门氏菌的最低检出量为6.2 CFU/mL;对24份样品的3次检测结果与国标法完全一致。证明建立的针对fimY蛋白基因的LAMP检测方法操作简便,特异性强,敏感性高,适用于现场检测。
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| 关 键 词： | 沙门氏菌 饲料 环介导等温扩增 快速诊断 |
| 收稿时间： | 2011-05-16 |
Development of a Loop-Mediated Isothermal Amplication Method for Rapid Detection of Salmonella in Animal Food |
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| SUN Yuan-yuan
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ZHAO Peng.Development of a Loop-Mediated Isothermal Amplication Method for Rapid Detection of Salmonella in Animal Food[J].China Animal Husbandry & Veterinary Medicine,2012,39(1):32-36. |
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| Authors: | SUN Yuan-yuan ZHAO Peng |
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| Institution: | Zhenjiang Entry-exit Inspection and Quarantine Bureau, Zhenjiang 212008, China |
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| Abstract: | According to the highly conserved fimY gene of Salmonella,reported in the GenBank,by primer explorer V4 softward,one set primer including four primers,were designed and the method for detecting the Salmonella with LAMP was developed after optimizing the reaction conditions.The results showed that the detection limit of the LAMP assay was 6.2 CFU/mL which was 100-fold higher than that of PCR.The test results of 24 samples by LAMP method was fully consistent with these by the national standard method.The results showed the LAMP method is more reliable and convenient for rapid,sensitive and specific diagnosis of Salmonella. |
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| Keywords: | Salmonella animal feed LAMP rapid detection |
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