Determination of the invA gene of Salmonella using surface plasmon resonance along with streptavidin aptamer amplification

被引:34
|
作者
Lei, Pinhua [1 ]
Tang, Hua [2 ]
Ding, Shijia [1 ]
Ding, Xiaojuan [1 ]
Zhu, Dan [1 ]
Shen, Bo [1 ]
Cheng, Quan [3 ]
Yan, Yurong [1 ]
机构
[1] Chongqing Med Univ, Coll Lab Med, Minist Educ, Key Lab Clin Lab Diagnost, Chongqing 400016, Peoples R China
[2] Chongqing Med Univ, Affiliated Hosp 2, Minist Educ, Key Lab Mol Biol Infect Dis, Chongqing 400016, Peoples R China
[3] Univ Calif Riverside, Dept Chem, Riverside, CA 92521 USA
基金
中国国家自然科学基金;
关键词
InvA; Salmonella; Streptavidin aptamer; Surface plasmon resonance; Asymmetric polymerase chain reaction; SELECTIVE ENRICHMENT BROTH; POLYMERASE-CHAIN-REACTION; SENSITIVE DETECTION; RAPID DETECTION; LABEL-FREE; PCR; ASSAY; TYPHIMURIUM; GRAPHENE; SENSOR;
D O I
10.1007/s00604-014-1330-6
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
We have developed a sensitive method for the determination of Salmonella by integrating a streptavidinylated aptamer (SA-aptamer) as a signal amplification unit along with a modified asymmetric polymerase chain reaction (PCR) technique into the surface of an SPR sensor chip. The gold film of the sensor was first modified with a thiolated probe, and the target sequence and SA-aptamer were then induced to form a sandwich structure. If SA is added, the SA-aptamer forms a complex with SA which will amplify the signal. Under optimal conditions, this sensing scheme has a linear response in the 50 pM to 200 nM range, and the lower detection limit is 20 pM (for a synthetic target sequence). This strategy was successfully applied to the determination of Salmonella bacteria at levels as low as 60 CFU mL(-1). This biosensor is sensitive, selective and highly stable. These features make this strategy a promising and powerful screening tool to detect pathogens in food, and in clinical and environmental samples.
引用
收藏
页码:289 / 296
页数:8
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