运用压电石英晶体生物传感器检测产气荚膜梭菌α毒素的实验研究  

Detecting Clostridium perfringens Alpha-toxin by Piezoelectric Quartz Crystal Biosensor

作　　者：赵渝徽[1] 陈鸣[1] 黄庆[1] 姚春艳[1] 闫慧慧[1] 匡红[1] 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

出　　处：《中华医院感染学杂志》2008年第3期320-323,共4页Chinese Journal of Nosocomiology

基　　金：国家863计划重大专项(2002AARZ2023);国家自然科学基金面上项目(3040010730400420);军队"十五"课题(01MA18001LO65);重庆市攻关课题(2004-20-12)

摘　　要：目的研究运用压电石英晶体生物传感器对产气荚膜梭菌α毒素(CPα)进行检测的方法和反应条件。方法按照碱基配对的原则,设计特异的产气荚膜梭菌α毒素基因寡核苷酸探针并利用巯基自组装技术固定在石英晶体上,运用压电石英晶体生物传感器的"质量效应"对聚合酶链反应(PCR)扩增得到的产气荚膜梭菌α毒素靶基因进行杂交检测,并探讨不同pH值和不同离子强度的缓冲液对检测的影响。结果压电石英晶体生物传感器成功检测出产气荚膜梭菌α毒素;pH7.6的缓冲液和0.64mol/L的NaCl溶液最适合于检测。结论压电石英晶体生物传感器能够有效地检测产气荚膜梭菌α毒素,有望对临床上开展毒素诊断提供一种新的手段。OBJECTIVE To search a method for identifying Clostridium perfringens alpha-toxin （CPα） by piezoelectric quartz crystal biosensor. METHODS A specific oligonucleotide probe for CPα gene according to the base pairing principle was designed and immobilized on the surface of quartz crystal with thiol self-assembly technique. Then CPα target gene obtained by PCR was detected by using the piezoelectric quartz crystal biosensor with mass effect. And the influence of different pH values and different ionic strength for detection were explored. RESULTS CPα was successfully detected by the piezoelectric quartz crystal biosensor. And the optimal conditions for detecting CPα were pH7.6 and 0.64mol/L NaCl solution. CONCLUSIONS The CPα could be detected efficiently by piezoelectric quartz crystal biosensor. Such method can provide a new modality for toxin diagnosis in clinic.

关 键 词：产气荚膜梭菌α毒素 基因 压电石英晶体生物传感器 寡核苷酸探针 

分 类 号：R378.8[医药卫生—病原生物学]