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BMC Infectious Diseases

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Open Access 01-12-2018 | Research article

Identification of nine cryptic species of Candida albicans, C. glabrata, and C. parapsilosis complexes using one-step multiplex PCR

Authors: Amir Arastehfar, Wenjie Fang, Weihua Pan, Wanqing Liao, Liang Yan, Teun Boekhout

Published in: BMC Infectious Diseases | Issue 1/2018

Abstract

Background

Candida albicans, Candida glabrata, and Candida parapsilosis are three prevalent causes of candidiasis, worldwide. These species are considered as nine medically important complex species. Limited knowledge about these newly recognized species prompted us to develop a one-step, multiplex PCR to detect and identify them in clinical settings.

Methods

Primers targeting Hyphal Wall Protein I gene for the C. albicans, C. dubliniensis, C. africana, Intergenic Spacer for the C. glabrata, C. nivariensis, C. bracarensis, and Intein and ITS rDNA for the C. parapsilosis, C. orthopsilosis, and C. metapsilosis were designed. Using 168 CBS reference strains and 280 clinical isolates, the specificity and reproducibility of the developed assay were evaluated.

Results

Our developed assay successfully identified and distinguished all the nine species. No cross-reaction with closely- and distantly-related yeast species, Aspergillus species and human DNA was observed, resulting in 100% specificity. The ambiguous results obtained by MALDI-TOF for C. albicans and C. africana were corrected by our 9-plex PCR assay. This assay identified all the cryptic complex species from two test sets from Iran and China, correctly.

Conclusions

Our developed multiplex assay is accurate, specific, cost/time-saving, and works without the tedious DNA extraction steps. It could be integrated into routine clinical laboratories and as a reliable identification tool and has the potential to be implemented into epidemiological studies to broaden the limited knowledge of cryptic species complexes.

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Title: Identification of nine cryptic species of Candida albicans, C. glabrata, and C. parapsilosis complexes using one-step multiplex PCR
Authors: Amir Arastehfar
Wenjie Fang
Weihua Pan
Wanqing Liao
Liang Yan
Teun Boekhout
Publication date: 01-12-2018
Publisher: BioMed Central
Published in: BMC Infectious Diseases / Issue 1/2018
Electronic ISSN: 1471-2334
DOI: https://doi.org/10.1186/s12879-018-3381-5

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Background

Methods

Results

Conclusions

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