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

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. Herei...

Full description

Bibliographic Details
Main Authors: Harmal, Nabil S., Khodavandi, Alireza, Alshawsh, Mohammed A., Jamal, Farida Fatema @ Farida, Sekawi, Zamberi, Ng, Kee Peng, Chong, Pei Pei
Format: Article
Language:English
Published: Academic Journals 2013
Online Access:http://psasir.upm.edu.my/id/eprint/29824/1/29824.pdf
_version_ 1825947587170009088
author Harmal, Nabil S.
Khodavandi, Alireza
Alshawsh, Mohammed A.
Jamal, Farida Fatema @ Farida
Sekawi, Zamberi
Ng, Kee Peng
Chong, Pei Pei
author_facet Harmal, Nabil S.
Khodavandi, Alireza
Alshawsh, Mohammed A.
Jamal, Farida Fatema @ Farida
Sekawi, Zamberi
Ng, Kee Peng
Chong, Pei Pei
author_sort Harmal, Nabil S.
collection UPM
description 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 albicans, Candida parapsilosis, Candida 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.
first_indexed 2024-03-06T08:15:40Z
format Article
id upm.eprints-29824
institution Universiti Putra Malaysia
language English
last_indexed 2024-03-06T08:15:40Z
publishDate 2013
publisher Academic Journals
record_format dspace
spelling upm.eprints-298242016-03-28T02:40:27Z http://psasir.upm.edu.my/id/eprint/29824/ Identification and differentiation of Candida species using specific polymerase chain reaction (PCR) amplification of the phospholipase B gene Harmal, Nabil S. Khodavandi, Alireza Alshawsh, Mohammed A. Jamal, Farida Fatema @ Farida Sekawi, Zamberi Ng, Kee Peng Chong, Pei Pei 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 albicans, Candida parapsilosis, Candida 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. Academic Journals 2013-05 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/29824/1/29824.pdf Harmal, Nabil S. and Khodavandi, Alireza and Alshawsh, Mohammed A. and Jamal, Farida Fatema @ Farida and Sekawi, Zamberi and Ng, Kee Peng and Chong, Pei Pei (2013) Identification and differentiation of Candida species using specific polymerase chain reaction (PCR) amplification of the phospholipase B gene. African Journal of Microbiology Research, 7 (20). art. no. C95F54C12002. pp. 2159-2166. ISSN 1996-0808 http://www.academicjournals.org/journal/AJMR/article-abstract/C95F54C12002
spellingShingle Harmal, Nabil S.
Khodavandi, Alireza
Alshawsh, Mohammed A.
Jamal, Farida Fatema @ Farida
Sekawi, Zamberi
Ng, Kee Peng
Chong, Pei Pei
Identification and differentiation of Candida species using specific polymerase chain reaction (PCR) amplification of the phospholipase B gene
title Identification and differentiation of Candida species using specific polymerase chain reaction (PCR) amplification of the phospholipase B gene
title_full Identification and differentiation of Candida species using specific polymerase chain reaction (PCR) amplification of the phospholipase B gene
title_fullStr Identification and differentiation of Candida species using specific polymerase chain reaction (PCR) amplification of the phospholipase B gene
title_full_unstemmed Identification and differentiation of Candida species using specific polymerase chain reaction (PCR) amplification of the phospholipase B gene
title_short Identification and differentiation of Candida species using specific polymerase chain reaction (PCR) amplification of the phospholipase B gene
title_sort identification and differentiation of candida species using specific polymerase chain reaction pcr amplification of the phospholipase b gene
url http://psasir.upm.edu.my/id/eprint/29824/1/29824.pdf
work_keys_str_mv AT harmalnabils identificationanddifferentiationofcandidaspeciesusingspecificpolymerasechainreactionpcramplificationofthephospholipasebgene
AT khodavandialireza identificationanddifferentiationofcandidaspeciesusingspecificpolymerasechainreactionpcramplificationofthephospholipasebgene
AT alshawshmohammeda identificationanddifferentiationofcandidaspeciesusingspecificpolymerasechainreactionpcramplificationofthephospholipasebgene
AT jamalfaridafatemafarida identificationanddifferentiationofcandidaspeciesusingspecificpolymerasechainreactionpcramplificationofthephospholipasebgene
AT sekawizamberi identificationanddifferentiationofcandidaspeciesusingspecificpolymerasechainreactionpcramplificationofthephospholipasebgene
AT ngkeepeng identificationanddifferentiationofcandidaspeciesusingspecificpolymerasechainreactionpcramplificationofthephospholipasebgene
AT chongpeipei identificationanddifferentiationofcandidaspeciesusingspecificpolymerasechainreactionpcramplificationofthephospholipasebgene