Objective <p><i>V. cholerae</i> is a Gram-negative bacterium that is transmitted to humans through ingestion of contaminated water and food. Rapid and accurate diagnosis of this bacterium is very important. So, the aim of this study was to design a biosensor using the plasmonic property of gold nanoparticles to detect <i>V. cholerae</i>.</p> Materials and methods <p>To make the biosensor, gold nanoparticles were first synthesized on grain particles according to the growth protocol. Nanostructures were evaluated and validated by UV-Vis spectroscopy, zeta potential, FT-IR and TEM. TA cloning of the target gene was performed and the recombinant plasmid was prepared. Also, the limit of detection (LOD), sensitivity, and specificity of the biosensor were determined and compared with real-time PCR and PCR assays.</p> Results <p>UV-Vis and TEM evaluations showed that nanoparticles were made in the form of rods. Based on the recombinant plasmid used, the LODs for biosensor, real-time PCR, and PCR were 1 × 10<sup>−8</sup>, 1 × 10<sup>−8</sup>, and 1 × 10<sup>−7</sup>&#xa0;ng of the target gene, respectively. Also, the specificity and sensitivity of all tests were 100%.</p> Conclusions <p>The biosensor significantly reduces the running time compared to the other two methods. Due to its high specificity and sensitivity, high speed, and ease of use, the biosensor is suitable for screening and diagnosis of <i>V. cholerae</i> and will be able to become a useful tool in clinical laboratories.</p>

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Fabrication of a sensitive surface plasmon resonance-based nanorod-shaped gold nanobiosensor for the specific detection of Vibrio cholerae

  • Samaneh Zadeh-Hadad,
  • Bita Bakhshi,
  • Tahereh Tohodi-Moghadam,
  • Saeed Shams

摘要

Objective

V. cholerae is a Gram-negative bacterium that is transmitted to humans through ingestion of contaminated water and food. Rapid and accurate diagnosis of this bacterium is very important. So, the aim of this study was to design a biosensor using the plasmonic property of gold nanoparticles to detect V. cholerae.

Materials and methods

To make the biosensor, gold nanoparticles were first synthesized on grain particles according to the growth protocol. Nanostructures were evaluated and validated by UV-Vis spectroscopy, zeta potential, FT-IR and TEM. TA cloning of the target gene was performed and the recombinant plasmid was prepared. Also, the limit of detection (LOD), sensitivity, and specificity of the biosensor were determined and compared with real-time PCR and PCR assays.

Results

UV-Vis and TEM evaluations showed that nanoparticles were made in the form of rods. Based on the recombinant plasmid used, the LODs for biosensor, real-time PCR, and PCR were 1 × 10−8, 1 × 10−8, and 1 × 10−7 ng of the target gene, respectively. Also, the specificity and sensitivity of all tests were 100%.

Conclusions

The biosensor significantly reduces the running time compared to the other two methods. Due to its high specificity and sensitivity, high speed, and ease of use, the biosensor is suitable for screening and diagnosis of V. cholerae and will be able to become a useful tool in clinical laboratories.