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目的建立金黄色葡萄球菌实时荧光PCR反应体系,用于消毒监测的快速检测。方法利用金黄色葡萄球菌耐热核酸酶nuc基因高保守序列,建立实时荧光PCR体系,评价其在消毒监测中的应用效果。结果建立的金黄色葡萄球菌实时荧光PCR反应体系检测时限短,全程1.5 h即可完成检测。该反应体系与其他细菌无交叉反应,重组阳性质粒和纯菌培养物检测下限分别为10拷贝/反应和35 cfu/ml。在1 880份消毒监测中所得阳性结果与细菌培养法结果一致。结论所建立的金黄色葡萄球菌nuc基因实时荧光PCR反应体系能准确检出标本中金黄色葡萄球菌,该方法快速、特异、灵敏。
Objective To establish a real-time fluorescence PCR reaction system of Staphylococcus aureus for the rapid detection of disinfection monitoring. Methods The high conserved sequence of nuc gene of heat-resistant nuclease of Staphylococcus aureus was used to establish a real-time fluorescence PCR system to evaluate its application in the disinfection monitoring. Results The detection time of S. aureus real-time fluorescence PCR reaction system was short, and the test was completed within 1.5 hours. The reaction system did not cross-react with other bacteria. The detection limits of the recombinant plasmid and pure culture were 10 copies / reaction and 35 cfu / ml, respectively. The positive results obtained in 1880 sterilization tests were consistent with the bacterial culture results. Conclusion The established real-time fluorescence PCR reaction system of staphylococcus aureus nuc gene can accurately detect Staphylococcus aureus in the sample. The method is rapid, specific and sensitive.