A novel electrochemical sensing strategy for rapid and ultrasensitive detection of Salmonella by rolling circle amplification and DNA-AuNPs probe

被引:77
|
作者
Zhu, Dan [1 ]
Yan, Yurong [1 ]
Lei, Pinhua [1 ]
Shen, Bo [1 ]
Cheng, Wei [1 ,3 ]
Ju, Huangxian [1 ,2 ]
Ding, Shijia [1 ]
机构
[1] Chongqing Med Univ, Coll Lab Med, Minist Educ, Key Lab Clin Lab Diagnost, Chongqing 400016, Peoples R China
[2] Nanjing Univ, Dept Chem, State Key Lab Analyt Chem Life Sci, Nanjing 210093, Jiangsu, Peoples R China
[3] Chongqing Med Univ, Affiliated Hosp 1, Ctr Clin Mol Med Detect, Chongqing 400016, Peoples R China
基金
中国国家自然科学基金;
关键词
Electrochemical biosensor; InvA gene; Salmonella; Rolling circle amplification; Gold nanoparticles; HIGH-SENSITIVE DETECTION; LABEL-FREE; GOLD NANOPARTICLES; TYPHIMURIUM; BACTERIA; APTASENSOR; BIOSENSOR; SEQUENCE; PCR;
D O I
10.1016/j.aca.2014.07.024
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A novel electrochemical sensing strategy was developed for ultrasensitive and rapid detection of Salmonella by combining the rolling circle amplification with DNA-AuNPs probe. The target DNA could be specifically captured by probe 1 on the sensing interface. Then the circularization mixture was added to form a typical sandwich structure. In the presence of dNTPs and phi29 DNA polymerase, the RCA was initiated to produce micrometer-long single-strand DNA. Finally, the detection probe (DNA-AuNPs) could recognize RCA product to produce enzymatic electrochemical signal. Under optimal conditions, the calibration curve of synthetic target DNA had good linearity from 10 aM to 10 pM with a detection limit of 6.76 aM (S/N = 3). The developed method had been successfully applied to detect Salmonella as low as 6 CFU mL(-1) in real milk sample. This proposed strategy showed great potential for clinical diagnosis, food safety and environmental monitoring. (C) 2014 Elsevier B.V. All rights reserved.
引用
收藏
页码:44 / 50
页数:7
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