| 金黄色葡萄球菌反向线性杂交探针检测方法的建立 |
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| 引用本文： | 韦莉,魏立雯,赖国旗,谭毅,张文露,潘永全. 金黄色葡萄球菌反向线性杂交探针检测方法的建立[J]. 中国实验动物学报, 2012, 0(6): 78-81,105 |
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| 作者姓名： | 韦莉 魏立雯 赖国旗 谭毅 张文露 潘永全 |
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| 作者单位： | 重庆医科大学实验动物中心;感染性疾病分子生物学重点实验室 |
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| 基金项目： | 重庆市科委科技创新能力建设项目《重庆市啮齿类实验动物工程技术研究中心》(CSTC,2010CB5013) |
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| 摘 要： | 目的建立一种检测金黄色葡萄球菌的简单、快速、灵敏、准确的方法。方法根据金黄色葡萄球菌的耐热核酸酶nuc基因,设计一对通用引物及两条特异性探针,用生物素标记通用引物的5＇端,将两条特异性探针固定于硝酸纤维膜上,使PCR产物与探针杂交。结果建立的反向线性杂交探针方法,其检测限为2 ng/μL,检测特异性和准确性均为100%。结论建立的反向线性杂交检测方法具有较高的敏感性和特异性,可用于实验动物金黄色葡萄球菌的快速检测。
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| 关 键 词： | 金黄色葡萄球菌 探针 反向线性杂交 |
Establishment of a detection technique of Staphylococcus aureus with reverse linear hybridization probes |
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| WEI Li,WEI Li-wen,LAI Guo-qi,TAN Yi,ZHANG Wen-lu,PAN Yong-quan. Establishment of a detection technique of Staphylococcus aureus with reverse linear hybridization probes[J]. Acta Laboratorium Animalis Scientia Sinica, 2012, 0(6): 78-81,105 |
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| Authors: | WEI Li WEI Li-wen LAI Guo-qi TAN Yi ZHANG Wen-lu PAN Yong-quan |
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| Affiliation: | 1(1.Experimental Animal Center,Chongqing Medical University,Chongqing 400016,China; 2.Key Laboratory of Molecular Biology of Infectious Diseases,Chongqing 400016) |
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| Abstract: | Objective To establish a simple, rapid, sensitive and accurate method for detection of Staphylococcus aureus （SA）. Methods According to SA thermostable nuclease nuc gene, a pair of universal primers and two specific probes were designed. 5＇ end of the universal primers were biotinylated. The probes were fixed on the nitrocellulose,then the probes were hybridized with PCR products. Results SA DNA concentration of 2 ng/μL could be detected by reverse linear hybrid method. The specificity and accuracy were 100%. Conclusion The reverse linear hybrid method has a high sensitivity and specificity. It can be used for rapid detection of SA. |
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| Keywords: | Staphylococcus aureus Probes Reverse linear hybridization |
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