通用引物pcr扩增创伤感染细菌16s-23s rdna间 - 第三军医大学学报.docVIP

纳米金颗粒质量放大压电DNA传感器频移信号的研究 蔡 晋1，王 珏1，夏 季2，罗 阳1，黄君富1，张晓莉1，府伟灵1 （第三军医大学：1西南医院检验科，重庆 400038；2大坪医院野战外科研究所检验科，重庆 400042） Study on gold nanoparticles mass amplification of signals in frequency shifts based piezoelectric DNA sensor CAI Jin1, WANG Jue1, XIA Ji2, LUO Yang1, HUANG Jun-fu1, ZHANG Xiao-li1, FU Wei-ling1 (Third Military Medical University: 1Department of Clinical Laboratory, Southwest Hospital, Chongqing 400038; 2Department of Clinical Laboratory, Daping Hospital, Chongqing 400042, China) Abstract：Objective To examine the feasibility of gold nanoparticles mass amplification of signals in frequency shifts based piezoelectric DNA sensor for improving the sensitivity of trace DNA detection. Methods The 5 nm gold nanoparticles were indirectly labled with target DNA to be measured after hybridization to increase mass loading on the suface of gold coated quartz crystal due to their relatively large mass compared to DNA molecule. Results It showed that the indirect labled gold nanoparticles significantly amplifyed the signals in frequency shifts of piezoelectric DNA sensor(P0.01). As low as 10-12 mol/L synthesized oligonucleotides could be detected by the sensor. Conclusion The use of gold nanoparticles effectively amplify the signals in frequency shifts resulted in the improved detection limit for synthesized oligonucleotides, which has lay a solid foundation for target DNA detection in next studies. Key words: gold nanoparticle; amplification; piezoelectric DNA sensor; frequency shift 压电DN传感器是近年发展起来的一种新的基因检测手段，与常规的核酸检测方法相比，压电DNA传感器具有液相杂交、实时检测、灵敏度高、特异性强、成本低、无污染、便于携带等特点而倍受人们青睐。纳米技术与生物传感技术相结合是DNA传感器发展的一大趋势。纳米金颗粒具有很好的生物相容性，易与DNA分子结合。此外，由于纳米金颗粒的质量远远大于DNA分子的质量，纳米金颗粒通过标记与DNA杂交体系结合后，即可增加石英晶振金膜表面的质量负载，从而降低响应频率，放大频移信号，提高传感器的检测灵敏度[1]。本研究以纳米金颗粒为DNA标记物，以探讨其放大压电DNA传感器的检测信号可行性。 1 材料与方法 1.1 实验材料 1.1.1 主要试剂纳米金颗粒溶液（5 nm粒径, 链亲和素标记, 90%, 体积比, TBS缓冲液稀释），购自美国KPL公司；牛血清白蛋白溶液（2% BSA, 质量体积比, TBS缓冲液溶解），新疆生化厂生产；无水乙醇，重庆川东化工（集团）有限公司生产；Piranha溶液（30%, 体积比, H2O2 : 浓H2SO4= 1:3），TBS缓冲液（含Tris-H