African Journal of
Microbiology Research

  • Abbreviation: Afr. J. Microbiol. Res.
  • Language: English
  • ISSN: 1996-0808
  • DOI: 10.5897/AJMR
  • Start Year: 2007
  • Published Articles: 5017

Full Length Research Paper

Identification and differentiation of Candida species using specific polymerase chain reaction (PCR) amplification of the phospholipase B gene

Nabil .S. Harmal1,2, Alireza Khodavandi3, Mohammed .A. Alshawsh4, Farida Jamal2, Zamberi Sekawi2, Ng Kee Peng5 and Pei Pei Chong6*
1Department of Medical Microbiology, Faculty of Medicine and Health Sciences, Sana’a University, Sana’a, Yemen. 2Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health Sciences, University Putra Malaysia, 43400 Selangor, Malaysia. 3Department of Basic Sciences, Gachsaran Branch, Islamic Azad University, Gachsaran, Iran. 4Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia. 5Department of Medical Microbiology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia. 6Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, University Putra Malaysia, 43400 Selangor, Malaysia.
Email: [email protected]

  •  Accepted: 19 April 2013
  •  Published: 14 May 2013

Abstract

Candida species are the major cause of mortality in both immunocompromised and critically ill patients. Therefore, the early diagnosis and differentiation of Candida species isolated from clinical samples is principally important due to their inherently variable antifungal resistance pattern. Herein, rapid and species-specific polymerase chain reaction (PCR)-based molecular method was developed for the identification of the four species of Candida most commonly isolated from clinical specimens, namely Candida albicansCandida parapsilosisCandida glabrata and Candida tropicalis. The developed method targeted the phospholipase B gene (PLB) as a novel target. We determined the sequences of this gene in previous work. The primers designed achieved highly specific identification of the selected species using simplex PCR assay, which were confirmed by sequencing. There was no cross-amplification of other Candida species nor other fungal organisms tested. The simplex PCR assay yielded detection limits of 1 to 10 cells/ml and 10 fg/µl DNA. These results showed that the PLB gene provides a novel target that could be used for the identification and detection of medically important Candida species from the clinical samples.

 

Key words: Candida species, identification, polymerase chain reaction (PCR), phospholipase B (PLB) gene.

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