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Highly specific and rapid molecular detection of Candida glabrata in clinical samples

  • Bacterial, Fungal and Virus Molecular Biology - Research Paper
  • Published:
Brazilian Journal of Microbiology Aims and scope Submit manuscript

Abstract

The most common nosocomial fungal infections are caused by several species of Candida, of which Candida glabrata is the second most frequently isolated species from bloodstream infections. C. glabrata displays relatively high minimal inhibitory concentration values (MIC) to the antifungal fluconazole and is associated with high mortality rates. To decrease mortality rates, the appropriate treatment must be administered promptly. C. glabrata contains in its genome several non-identical copies of species-specific sequences. We designed three pairs of C. glabrata-specific primers for endpoint PCR amplification that align to these species-specific sequences and amplify the different copies in the genome. Using these primers, we developed a fast, sensitive, inexpensive, and highly specific PCR-based method to positively detect C. glabrata DNA in a concentration-dependent manner from mixes of purified genomic DNA of several Candida species, as well as from hemocultures and urine clinical samples. This tool can be used for positive identification of C. glabrata in the clinic.

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Data availability

All plasmids and yeast strains are available upon request.

The oligonucleotide pairs Cg1, Cg2, and Cg3 described in this work were designed by A. D. L. P. and have been included in patents MX352246 B and EP2410052 B1.

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Acknowledgements

The authors thank Dr. Lina Riego, Dr. Héctor Mora, and Dr. Celia Pais for providing reference strains and Dr. Areli Martínez, Dr. José Sifuentes, Dr. Pedro Torres from Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán: INCMNSZ, and Dr. Javier Araujo from Hospital Central Ignacio Morones Prieto for providing Candida sp. clinical isolates and blood cultures.

We thank Mayra Cuéllar-Cruz for technical support in the initial experiments.

The authors also thank Verónica Zárate and LANBAMA-IPICYT for sample sequencing.

Funding

This work was supported by Consejo Nacional de Ciencia y Tecnología (CONACyT) Grants No. CB 2005–01-48304 and No. CB 2014–239629 to I.C. and Fondo Sectorial de Salud-CONACyT to A. D. L. P. No. 2005–13927.

O.H.-C.; C.H.-H.; and M.S.H.-B. were supported by CONACyT fellowship nos. 611321, 279,071, and 932,042 respectively. B.E.G.-G. was supported by a postdoctoral fellowship from CONACyT No. 205158 and G.H.-H. received a CEN BES fellowship by Secretaría de Educación Pública and a fellowship from IPICYT.

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Authors and Affiliations

  1. IPICYT, División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica, AC, Camino a la Presa San José No. 2055, Col. Lomas 4, 78216, San Luis Potosí, Mexico

    Oscar Hernández-Carreón, Cesia Hernández-Howell, Grecia Hernández-Hernández, M. Selene Herrera-Basurto, Blanca E. González-Gómez, Guadalupe Gutiérrez-Escobedo, Norma I. García-Calderón, Daniel Barrón-Pastor, Alejandro De Las Peñas & Irene Castaño

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  1. Oscar Hernández-Carreón
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Contributions

C. H. H.; G. H. H.; and B. E. G. G. were involved in the optimization of PCR conditions. O. H. C.; C. H. H.; G. H. H.; M. S. H. B.; and B. E. G. G. performed PCR identification of Candida isolates, hemocultures, and urine samples and API® ID 32C validation experiments. G. G. E. planned and executed experiments. O. H. C. and C. H.-H. determined minimal amount of DNA detected. N. I. G. G. and D. B. P. wrote and followed up the patent applications. A. D. L. P. and I. C. were involved in conceptualization, design of the experiments, and design of the species-specific oligonucleotide pairs. I. C. wrote the manuscript and all authors were involved in reviewing and editing of the manuscript.

Corresponding author

Correspondence to Irene Castaño.

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Hernández-Carreón, O., Hernández-Howell, C., Hernández-Hernández, G. et al. Highly specific and rapid molecular detection of Candida glabrata in clinical samples. Braz J Microbiol 52, 1733–1744 (2021). https://doi.org/10.1007/s42770-021-00584-2

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  • DOI: https://doi.org/10.1007/s42770-021-00584-2

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