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A Nanodiagnostic Colorimetric Assay for 18S rRNA of Leishmania Pathogens using Nucleic Acid Sequence-Based Amplification and Gold Nanorods

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Abstract

Background and Objective

We describe here a nanodiagnostic colorimetric assay for 18S rRNA of Leishmania pathogens that uses nucleic acid sequence-based amplification (NASBA) and gold nanorods (GNRs).

Methods

NASBA primers targeting 18S rRNA were used for amplification of RNA in an isothermal process. The electrostatic interactions between the phosphate groups of the RNA amplicons and the cetyl trimethylammonium bromide layer on the GNRs resulting in their aggregation. This phenomenon changes the color of the GNR solution from red to purple.

Results

Our data showed 100 % sensitivity and 80 % specificity with the colorimetric assay compared with results using reverse transcription polymerase chain reaction.

Conclusion

The nanodiagnostic method we describe simplifies the detection of NASBA amplicons without the need for gel electrophoresis.

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Acknowledgments

This study was supported by the Vice-Chancellor in Research and Technology Affairs, Golestan University of Medical Sciences. The Iran Nanotechnology Initiative (INI) also supported partially this project. The authors thank Fatemeh Ghaffarifar, Fatemeh Mesgarian, Sareh Zhand, and Mohammad Shariati for their help in the laboratory experiments.

Author Contributions

P. Gill conceived and designed the experiments. A. Niazi and H. Nikbakht performed the experiments. P. Gill and O-N Jorjani analyzed the data. O-N. Jorjani and P. Gill contributed reagents/materials/analysis tools. A. Niazi and P. Gill wrote the paper.

Conflict of Interest

The authors declare that they have no conflict of interest that are directly relevant to the content of this article.

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Authors and Affiliations

  1. Department of Medical Biotechnology, Faculty of Advanced Medical Technologies, Golestan University of Medical Sciences, Gorgan, Iran

    Alireza Niazi & Hassan Nikbakht

  2. Department of Medical Laboratory Sciences, Faculty of Paramedicine, Golestan University of Medical Sciences, Gorgan, Iran

    Oghol-Niaz Jorjani

  3. Laboratory Science Research Center, Golestan University of Medical Sciences, Gorgan, Iran

    Oghol-Niaz Jorjani

  4. Department of Medical Nanotechnology, Faculty of Advanced Medical Technologies, Golestan University of Medical Sciences, Gorgan, Iran

    Pooria Gill

Authors
  1. Alireza Niazi
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  2. Oghol-Niaz Jorjani
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  3. Hassan Nikbakht
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  4. Pooria Gill
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Correspondence to Oghol-Niaz Jorjani or Pooria Gill.

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Niazi, A., Jorjani, ON., Nikbakht, H. et al. A Nanodiagnostic Colorimetric Assay for 18S rRNA of Leishmania Pathogens using Nucleic Acid Sequence-Based Amplification and Gold Nanorods. Mol Diagn Ther 17, 363–370 (2013). https://doi.org/10.1007/s40291-013-0044-5

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  • DOI: https://doi.org/10.1007/s40291-013-0044-5

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