基于自组装单分子层技术的压电dna传感器固定-第三军医大学学报.doc

核酸外切酶-压电石英DNA传感器检测金黄色葡萄球菌的实验研究( 黄君富，夏涵，黄庆，府伟灵（第三军医大学西南医院检验科，重庆 400038） 摘 要：目的 构建一种用于检测金黄色葡萄球菌核酸外切酶-压电石英DNA传感器新方法，同时对其重要影响因素进行优化探讨。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（p0.01），可以检测到1.0×104 CFU/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。 关键词：石英晶体微天平；生物传感器；λ核酸外切酶；金黄色葡萄球菌 Detection of Staphylococcus aureus using a lambda exonuclease based piezoelectric Quartz Crystal Microbalance DNA Sensor HUANG Jun-fu, XIA Han, HUANG Qing, FU Wei-ling (Southwest Hospital, the Third Military Medical University, Chongqing 400038，China) Abstract: Objective To develop a new type of piezoelectric quartz crystal microbalance DNA sensor based on lambda exonuclease. Methods After been treated with Lambda exonuclease, the PCR products were visualized by agarose gel electrophoresis with ethidium bromide, and then analyzed by a gel documentation system. The temperature and time of hybridization were optimized respectively. In addition, we detected the PCR products of Staphylococcus aureus with this piezoelectric quartz crystal microbalance DNA sensor system to assess its specificity and sensitivity. Results The optimum hybridization temperature was 35℃ and the optimum hybridization time was 60 min. The response performance of the lambda exonuclease based piezoelectric quartz crystal microbalance DNA sensor was better than that of common DNA sensor. The lowest detection limit of current quartz crystal microbalance system was 1.0×104 CFU/mL. Conclusion The lambda exonuclease based quartz crystal microbalance system we designed has the advantages of high efficiency hybridization, easy to operate, and good response performance. Thus it can be applied in Staphylococcus aureus infection detections. Key words: Quartz crystal microbalance; Biosensor; Lambda exonuclease; Staphylococcus aureus 金黄色葡萄球菌是人类化脓性感染的重要病原菌，毒力强，