Highly selective and sensitive detection of Staphylococcus aureus with gold nanoparticle-based core-shell nano biosensor

被引:34
|
作者
Shahbazi, Reza [1 ]
Salouti, Mojtaba [1 ]
Amini, Bahram [2 ]
Jalilvand, Ahmad [3 ]
Naderlou, Ebrahim [1 ]
Amini, Ali [2 ]
Shams, Arash [1 ]
机构
[1] Islamic Azad Univ, Zanjan Branch, Biol Res Ctr, Zanjan, Iran
[2] Zanjan Univ, Fac Sci, Dept Biol, Zanjan, Iran
[3] Zanjan Univ Med Sci, Dept Pathol, Pathol, Zanjan, Iran
关键词
S; aureus; Colorimetry; AuNPs; Protein A gene; MAGNETIC NANOPARTICLES; COLORIMETRIC SENSOR; CLINICAL-SAMPLES; GENE; PROBE;
D O I
10.1016/j.mcp.2018.07.004
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Staphylococcus aureus is a gram-positive and opportunistic pathogen that is one of the most common causes of nosocomial infections; therefore, its rapid diagnosis is important and valuable. Today, the use of nanoparticles is expanding due to their unique properties. The purpose of the present study is the determination of S. aureus by a colorimetric method based on gold nanoparticles (AuNPs). Firstly, S. aureus was cultured on both LB media (broth and agar) and their chromosomal DNA was extracted. Afterwards, primers and biosensor were designed based on Protein A sequence data in the gene bank. PCR assay was performed under optimal conditions and the PCR product was electrophoresed on 2-percent agarose gel. The synthesized biosensors were conjugated with AuNPs and, eventually, a single-stranded genome was added to the conjugated AuNPs and hybridization was performed. The results were evaluated based on color change detected by the naked eye, optical spectrophotometry, and transient electron microscopy. Finally, the sensitivity and specificity of the AuNP-biosensor were determined. The results of the present study showed a 390 bp band on the agarose electrophoresis gel, which confirmed the presence of Protein A genes on the chromosome of the bacteria. The PCR and calorimetric methods were compared with each other. The sensitivity of the PCR and colorimetric methods were 30 ng mu g(-1) and 10 ng mu L-1, respectively. The limit of detection (LOD) equaling 8.73 ng mu L-1 was determined and the specificity of the method was confirmed by the DNA of other bacteria. According to the results, the present method is rapid and sensitive in detecting S. aureus.
引用
收藏
页码:8 / 13
页数:6
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