新型漏声表面波生物传感器液相检测的实验研究-第三军医大学学报.docVIP

漏声表面波传感器生物信号放大系统的研究 王云霞1，张立群1，罗 阳1，丁 毅2，施建峰2，徐清华3，汪广杰1，曹 亮3，陈 鸣3(府伟灵1( 利用“RecA蛋白-互补单链DNA探针与bis-PNA/dsDNA复合物相结合，提高传感器的检测灵敏度。方法传感器表面先固定人乳头瘤病毒(HPV)的bis-PNA探针，加入HPV基因组DNA与之完全反应后，加入不同浓度“RecA蛋白-互补单链DNA”探针ATPγS提供能量。RecA蛋白及ATPγS的最佳结果 当RecA蛋白浓度为45 μg/mL，ATPγS浓度为2.5 mmol/L 时所引起的相位变化值最为显著。结论 “RecA蛋白-互补单链DNA”探针复合体与bis-PNA/dsDNA复合物相结合有效提高传感器的检测灵敏度。漏声表面波；传感器；；RecA蛋白灵敏度 The Study of Biological Signal Amplification System of Leaky Surface Acoustic Wave senser WANG Yun-xia, ZHANG Li-qun, LUO Yang, DING Yi, SHI Jian-feng, Xu Qing-hua，WANG Guang-jie, CAO Liang, CHEN Ming, FU Wei-ling (Department of Laboratory Medicine, Southwest Hospital, the Third Military Medical University, Chongqing 400038, China） Abstract: Objective　To improve the sensitivity of leaky surface acoustic wave bis- peptide nucleic acid biosensor by “RecA protein-complementary single strand DNA probe” complex as biological signal amplification system. Methods Bis-PNA probe for detecting HPV was immobilized on the surface of detection channel firstly. Bis-PNA probe was hybridized with corresponding target DNA complately, Then the different concentrations of the “RecA protein-complementary single strand DNA probe” was added to react with bis-PNA/dsDNA complex. Results The phase shift arrived maximum when the concentration of RecA protein was 45μg/mL and that of ATPγS was 2.5mmol/L. Conclusion The sensitivity of leaky surface acoustic wave biosensor is improved effectively by adding “RecA protein-complementary single strand DNA probe” complex. Key word：leaky surface acoustic wave; sensor; peptide nucleic acid; RecA protein; sensitivity 随着分子生物学的发展，已成为临床病原学早期诊断的重要手段之一漏声表面波(eaky surface acoustic wave，LSAW)[1-2]，针对待检靶DNA设计的检测探针可以提高传感器的特异性。与传统的DNA探针不同，本实验中我们设计合成了更稳定更特异的肽核酸(peptide nucleic acid, PNA)探针。PNA是人工合成的DNA模拟物，其中双体肽核酸(bis-PNA)更是以“链侵袭”的方式与双链DNA形成稳定性更高的四链体结构，可直接检测DNA靶序列[3]。为了进一步提高传感器检测的灵敏度，本研究将“RecA蛋白-互补单链DNA探针”引入到LSAW-bisPNA生物传感器检测系统，利用RecA蛋白可以促使同源DNA分子碱基配对1.1 实验材料 LSAW传感器的制作 由本室与中国电子科技集团公司第26研究所共同研制，设计为对端双谐振型传感器。即在同一