漏声表面波传感器生物信号放大系统的研究  

Study on biological signal amplification system of leaky surface acoustic wave sensor

作　　者：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] 

机构地区：[1]第三军医大学西南医院检验科,重庆400038 [2]中国电子科技集团公司第二十六研究所五室,重庆400060 [3]第三军医大学大坪医院野战外科研究所检验科,重庆400042

出　　处：《第三军医大学学报》2009年第11期1013-1016,共4页Journal of Third Military Medical University

基　　金：国家863探索导向性课题(2007AA02Z416);国家863计划重点项目(2007AA022005);国家自然科学基金(30400107)~~

摘　　要：目的研究利用"RecA蛋白-互补单链DNA"探针与bis-PNA/dsDNA复合物相结合,作为生物信号放大系统提高传感器检测灵敏度的可行性。方法漏声表面波传感器检测通道金膜表面先固定针对人乳头瘤病毒(HPV)的bis-PNA探针,加入HPV基因组DNA与之完全反应后,再加入不同浓度的"RecA蛋白-互补单链DNA"探针,在反应过程中加入ATPγS提供能量,分别探索优化RecA蛋白及ATPγS的最佳反应浓度。结果当RecA蛋白浓度为45μg/ml,所引起的相位变化值为(11.74±1.03)°,显著高于其他浓度组(P<0.01);在最佳RecA蛋白浓度下,当ATPγS浓度为2.5 mmol/L时,所引起的相位变化值为(10.71±0.73)°,显著高于其他ATPγS浓度组(P<0.01)。结论"RecA蛋白-互补单链DNA"探针复合体与bis-PNA/dsDNA复合物相结合可以有效提高传感器的检测灵敏度,并明显缩短检测时间。Objective To study the feasibility of combination of ＂RecA protein-complementary single strand DNA＂ probe and bis-PNA/dsDNA complexes as the biological signal amplification system for the improvement of the sensitivity of leaky surface acoustic wave bispeptide nucleic acid biosensor. Methods Bis- PNA probe for detecting human papilloma virus （HPV） was immobilized on the surface of gold membrane of the detection channel. After the probe was hybridized with the corresponding target DNA completely, different concentrations of ＂RecA protein-complementary single strand DNA probe＂ were added to react with bis-PNA/dsD- NA complexes. Results The phase shift value was （ 11.74 ± 1.03 ） deg at the concentration of 45 μg/ml of RecA protein, significantly different as compared with other concentrations （P 〈 0.01 ）. At the optimal concentration of RecA protein, the phase shift value was （ 10.71 ±0.73） deg at the concentration of 2.5 mmol/L of ATPγS, significantly different as compared with other concentrations （ P 〈 0.01 ）. Conclusion Combination of ＂RecA protein-complementary single strand DNA probe＂ and bis-PNA/dsDNA complexes contributes to ef- fective improvement of the sensitivity and significantly shortened detection time.

关 键 词：漏声表面波 传感器 肽核酸 RecA蛋白 灵敏度 

分 类 号：R318.04[医药卫生—生物医学工程] R394-33[医药卫生—基础医学]