Specific Identification of Candida Glabrata via Colorimetric Assay Based on Gold Nanoparticles

Document Type : Original Article (s)

Authors

1 MSc Student, Department of Parasitology and Mycology, School of Medicine AND Student Research Committee, Isfahan University of Medical Sciences, Isfahan, Iran

2 Assistant Professor, Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

3 Associate Professor, Biology Research Center, Islamic Azad University, Zanjan Branch, Zanjan, Iran

4 Assistant Professor, Department of Genetics, School of Sciences, Islamic Azad University, Zanjan Branch, Zanjan, Iran

Abstract

Background: Candida glabrata is one of the most common Candida species which is able to get resistant to antifungals. So, the rapid identification of it, seems to be necessary for early treatment of infections. The conventional methods for detection of Candida species are time-consuming and difficult. Today, using of nanoparticles with unique properties has been expanded. The purpose of this study was rapid identification of Candida glabrata using colorimetric assay based on gold nanoparticles.Methods: Candida glabrata was cultured in yeast extract peptone dextrose broth medium and DNA was extracted. The gene specific sequence (CAGL0M05005g) was determined from the Candida genetic information bank. For polymerase chain reaction (PCR), the primers and probe were designed via Oligo 7 software. The PCR product and probe was affected by denaturation and optimized binding temperatures. The gold nanoparticles were added and changed the color to be visual; by using ultraviolet-visible (UV-vis) spectrophotometer and transmission electron microscopy (TEM), they were confirmed. To evaluate the sensitivity of new method, different concentrations of PCR product were used and the results were analyzed using gel electrophoresis.Findings: The optimum temperatures of the primer binding to the DNA template and hybridization the probe was determined. After adding gold nanoparticles, the mixture solution color was changed from red to purple. The results were confirmed via spectroscopy and electron microscopy.Conclusion: The results indicated that in detection of Candida glabrata, the new method is faster than conventional methods and more sensitive than gel electrophoresis. 

Keywords

References

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Journal of Isfahan Medical School
Volume 33, Issue 325 - Serial Number 325
January and February 2015
Pages 231-241

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History

  • Receive Date: 01 February 2015
  • Revise Date: 01 November 2024
  • Accept Date: 06 April 2022

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