PCR技术在鼠金黄色葡萄球菌检测中的初步研究

根据已公布的金黄色葡萄球菌耐热核酸酶nuc基因的序列 ,设计并合成一对特异性的引物 ,利用PCR技术扩增nuc基因片段。对金黄色葡萄球菌和其他非金黄色葡萄球菌菌株抽提的DNA进行扩增。结果金黄色葡萄球菌PCR产物出现 6 6 8bp的特异性DNA扩增片段 ,而其他非金黄色葡萄球菌未出现扩增片段 ,证实了合成的引物对金黄色葡萄球菌具有特异性。将抽提的金黄色葡萄球菌DNA进行系列稀释 ,测定此PCR体系的敏感性。结果显示 ,该PCR体系能检出 3pg金黄色葡萄球菌DNA ,且从抽提DNA到PCR扩增及电泳结束仅需 4h。因此 ,研究所建立的扩增耐热核酸酶nuc基因检测鼠金黄色葡萄球菌的PCR方法 ,具有快速、可靠、敏感和特异的特点 ,可用于临床样品和金黄色葡萄球菌感染时的检测 ,适合应用于实验大小鼠的监测

The pilot study of polymerase chain reaction for detecting Staphylococcus aureus in rats and mice

This study aimed to establish a rapid method for detection of Staphylococcus aureus in laboratory rats and mice.According to the sequence of the nuc gene,encodeing the thermostable nuclease of Staphylococcus aureus,a pair of oligonucleotide primers were designed and synthesized to amplify a sequence of nuc gene by PCR.The PCR product was detected by agarose gelelectrophoresis analysis.The results indicated the PCR could detect as little as 3 pg Staphylococcus aureus DNA.The whole process of DNA extraction amplification and electrophoresis could be completed within 4 h.The PCR assay was a rapid,reliable,sensitive and specific method of detecting Staphylococcus aureus,which provides a useful tool for direct applification of clinical specimens,and may be useful for diagnosis of aureus staphylococcal infections in rats and mice.