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目的构建一种用于检测金黄色葡萄球菌的核酸外切酶-压电石英DNA传感器新方法,同时对其重要影响因素进行探讨。方法采用λ核酸外切酶处理金黄色葡萄球菌PCR产物,EB染色、电泳后于紫外灯下进行观察,对杂交温度、杂交时间和杂交响应性能等因素进行实验研究,进一步将构建的核酸外切酶-压电石英DNA传感器法用于金黄色葡萄球菌检测,并对其灵敏度和特异性进行研究。结果λ核酸外切酶-压电石英DNA传感器法最适杂交温度为35℃,最适杂交时间为60min,响应性能显著大于普通压电DNA传感器(P<0.01),可以检测到1.0×104CFU/ml的金黄色葡萄球菌,特异性好。结论利用核酸外切酶-压电石英DNA传感器检测金黄色葡萄球菌,具有杂交效率高、技术难度低、频率响应性能高等优点,有较好的应用前景。
OBJECTIVE: To construct a novel exonuclease - DNA piezoelectric sensor for detection of Staphylococcus aureus, and discuss its important influence factors. Methods The PCR products of Staphylococcus aureus were treated with λ exonuclease and stained with EB. After electrophoresis, the cells were observed under UV light. The temperature, hybridization time and hybridization response were studied. The constructed exonucleolytic The enzyme-piezoelectric quartz DNA sensor method was used for the detection of Staphylococcus aureus and its sensitivity and specificity were studied. Results The optimal exonuclease-piezoelectric quartz DNA sensor method was 35 ℃, the optimum hybridization time was 60min, the response performance was significantly higher than that of the ordinary piezoelectric DNA sensor (P <0.01), and 1.0 × 104CFU / ml of Staphylococcus aureus, specificity is good. Conclusion The detection of Staphylococcus aureus with exonuclease - piezoelectric quartz DNA sensor has the advantages of high hybridization efficiency, low technical difficulty and high frequency response, which has a good application prospect.