MRAB OXA-23基因检测电化学DNA传感器的构建及初步评价  

Establishment on electrochemical DNA sensor for MRAB OXA-23 gene determination

作　　者：郭宏波[1] 戴飘飘 黄华亮 郭彦伟[1] GUO Hongbo;DAI Piaopiao;HUANG Hualiang;GUO Yanwei(Department of Clinical Laboratory,Inner MongoliaBaogang Hospital,Baotou 014010,Inner Mongolia,China;Department of Clinical Laboratory, Taizhou Municipal Hospital,Taizhou 318000,Zhejiang,China)

机构地区：[1]内蒙古包钢医院检验科,内蒙古包头014010 [2]台州市立医院检验科,浙江台州318000

出　　处：《检验医学》2019年第4期346-350,共5页Laboratory Medicine

摘　　要：目的构建用于检测多重耐药鲍曼不动杆菌(MRAB)OXA-23突变基因的新型电化学DNA传感器。方法采用多分枝长针海胆形铂纳米树枝(Pt-LSSUs@PAA)修饰电化学DNA传感器界面,以六氨合钌(RuHex)为信号指示剂,通过差分脉冲伏安法检测传感器杂交前后与DNA骨架中带负电的磷酸基团结合的RuHex产生的电化学信号差异。用构建完成的电化学DNA传感器定量检测MRAB OXA-23突变基因。结果构建的电化学DNA传感器具有良好的稳定性和重现性,Pt-LSSUs@PAA的修饰能显著增大电极的有效面积,加快电极表面的电子传导速率,达到信号放大的目的。构建完成的电化学DNA传感器测定OXA-23浓度的线性范围为10 fmol/L～10 nmol/L,检测限为3.3 fmol/L(信噪比为3),且具有良好的特异性。结论构建的电化学DNA传感器可灵敏、特异地定量检测MRAB OXA-23基因。Objective To establish a new electrochemical DNA sensor for determining multidrug-resistant Acinetobacter baumannii(MRAB) OXA-23 gene. Methods Polyallylamine-functionalized Pt nanostructures with long-spined sea-urchin-like morphology(Pt-LSSUs@PAA) was used to modify electrochemical DNA sensor interface. RuHex was used as a signal indicator. Using the established electrochemical DNA sensor,the differential pulse voltammetry was used to determine the presence of negatively charged phosphorus-bound RuHex electrochemical signal differences generated by OXA-23 mutant gene to be determined quantitatively. Results The sensor had good stability and reproducibility. The modification of Pt-LSSUs@PAA can increase the effective area of electrode,speed up the electron conduction rate on the surface of electrode and achieve signal amplification. The linear range of OXA-23 was 10 fmol/L-10 nmol/L. The determination limit was 3.3 fmol/L(signal-to-noise ratio was 3),and the sensor had good specificity. Conclusions The established sensor can achieve sensitive and specific quantitative determination of OXA-23 mutant gene.

关 键 词：OXA-23基因 多重耐药鲍曼不动杆菌 多分枝长针海胆形铂纳米树枝 DNA传感器 

分 类 号：R446.1[医药卫生—诊断学]