摘要
基于荧光探针法建立一种快速、灵敏的金黄色葡萄球菌核酸检测方法.针对金黄色葡萄球菌nuc基因设计特异性引物和探针,制备质粒标准品和菌液核酸标准品,完成方法建立.然后,对该方法的特异性、敏感性、重复性、实用性等性能进行评价.结果显示,该方法特异性100％;质粒标准品敏感性为1 copy/μL;菌液标准品敏感性为1 000CFU/mL;重复性变异系数为0.69％;多种基质加标样品均能有效检出.成功建立了一种快速、灵敏的金黄色葡萄球菌核酸检测方法,可用于疑似金黄色葡萄球菌污染样品的快速核酸检测.
Abstract
To establish a rapid,sensitive fluorescent probe polymerase chain reaction(PCR)method for the detection of Staphylococcus aureus.Specific primers and probes were designed for the Staphylococcus aureus nuc gene,and the plasmid standard and pure culture standard were prepared,and the method was established.Then,the specificity,sensitivity,repeatabili-ty and practicability of the method were evaluated.As a result,the specificity of this method was 100％,the sensitivity of plasmid was 1 copy/μL,the sensitivity of pure culture level was 1 000 CFU/mL,the coefficient of variation of repeatability was 0.69％,various matrix spiked samples were effectively detected.A fast and sensitive method for nucleic acid detection has been successfully established,which can be used for rapid nucleic acid detection of suspected Staphylococcus aureus contam-inated samples.
维普
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